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Dental focal infections are extraoral manifestations caused by oral pathogens. Pathological oral conditions, such as periapical inflammation and periodontitis, can cause bacteremia. Dissemination of oral pathogens to nonoral sites can subsequently cause infections in extraoral tissues and organs. Cardiovascular infections and brain abscesses are the most common of these. The course of such infections can be lethal. In order to improve patient care, a closer collaboration between dental and medical caregivers is necessary.Dentale focale infecties zijn extraorale ontstekingen die worden veroorzaakt door orale pathogenen. Orale aandoeningen als periapicale ontsteking of parodontitis kunnen bacteriëmie veroorzaken. Disseminatie van orale pathogenen kan vervolgens leiden tot infecties in extraorale organen en weefsels. Hierbij komen cardiovasculaire infecties en hersenabcessen het meest frequent voor. Dergelijke infecties kunnen levensbedreigend verlopen. Om de patiëntenzorg te kunnen verbeteren is intensievere samenwerking tussen tandheelkundige en geneeskundige zorgverleners noodzakelijk.

Clinical microbiology now occupies an important place in periodontics and oral implant dentistry as a supplementary diagnostic tool and in planning treatment, particularly with respect to the rational use of antibiotics. This view is in line with the emphatic call by the World Health Organization and the European Union for the prudent use of antibiotics due to the global increase in resistance to antibiotics. Furthermore side effects may occur, such as the disturbance of the microbial intestinal and oral microflora, sometimes leading to serious pathological conditions. Hyposalivation following the use of antibiotics may lead to an oral environmental condition in which caries may develop faster than usual.Klinische microbiologie neemt inmiddels een belangrijke plaats in binnen de parodontologie en de orale implantologie als aanvullend diagnosticum en voor behandelplanning, vooral bij het rationeel gebruik van antibiotica. Deze zienswijze sluit aan bij de nadrukkelijke oproep van de World Health Organization en de Europese Unie om prudent om te gaan met antibiotica vanwege een wereldwijde toename in antibioticumresistentie. Daarnaast kunnen antibiotica bijwerkingen geven zoals verstoring van de darmflora en het orale milieu met soms ernstige pathologie tot gevolg. Hyposalivatie na antibioticumgebruik kan leiden tot een mondmilieu waarin cariës zich sneller kan ontwikkelen.

Porphyromonas gingivalis is one of the major oral pathogens implicated in the widespread inflammatory disorder periodontitis. Moreover, in recent years, P. gingivalis has been associated with the autoimmune disease rheumatoid arthritis. The peptidylarginine deiminase enzyme of P. gingivalis (PPAD) is a major virulence factor that catalyzes the citrullination of both bacterial and host proteins, potentially contributing to production of anticitrullinated protein antibodies. Considering that these antibodies are very specific for rheumatoid arthritis, PPAD appears to be a link between P. gingivalis, periodontitis, and the autoimmune disorder rheumatoid arthritis. PPAD was thus far considered unique among prokaryotes, with P. gingivalis being the only bacterium known to produce and secrete it. To challenge this hypothesis, we investigated the possible secretion of PPAD by 11 previously collected Porphyromonas isolates from a dog, 2 sheep, 3 cats, 4 monkeys, and a jaguar with periodontitis. Our analyses uncovered the presence of secreted PPAD homologues in 8 isolates that were identified as Porphyromonas gulae (from a dog, monkeys, and cats) and Porphyromonas loveana (from sheep). In all 3 PPAD-producing Porphyromonas species, the dominant form of the secreted PPAD was associated with outer membrane vesicles, while a minor fraction was soluble. Our results prove for the first time that the citrullinating PPAD exoenzyme is not unique to only 1 prokaryotic species. Instead, we show that PPAD is produced by at least 2 other oral pathogens.

De aanwijzingen voor een verband tussen systemische ziekten en parodontitis zijn het sterkst bij diabetes mellitus type 2 en hart- en vaatziekten. Er is sprake van een matig verband van parodontitis met zwangerschapscomplicaties en reumatoïde artritis. Parodontale behandeling draagt gemiddeld genomen bij aan een verlaging van de systemische ontsteking en een verbetering van de conditie van het bloedvatenstelsel. Bij diabetespatiënten kan parodontale behandeling ook een positief effect hebben op de metabole regulatie. Er is onvoldoende bewijs dat parodontale behandeling een positief effect kan hebben op afgeleide parameters van zwangerschapscomplicaties en reumatoïde artritis.

Treatment of infectious diseases is complex and success is dependent upon a number of factors. The proper prescription and correct dosage of antimicrobials are important elements in this respect. Incorrect prescription and insufficient dosage can result not only in treatment failure but also contribute to the development of antimicrobial resistance. In order to achieve correct antibiotic dosages it is necessary to gain insight in pharmacokinetic and pharmacodynamic principles. It is also vital to differentiate among various types of infections and to adjust antibiotic prescription to the clinical situation of the individual patient accordingly. To further optimise antibiotic policy collaboration between dental and medical professions is of major importance.De behandeling van infectieziekten is complex en het slagen ervan is van diverse factoren afhankelijk. Het juist voorschrijven en correct doseren van antibiotica is hierbij een belangrijk onderdeel. Als hierin tekort wordt geschoten dan kan dit niet alleen leiden tot therapiefalen, maar ook bijdragen aan resistentie-ontwikkeling. Voor het correct doseren van antibiotica is inzicht in farmacodynamische en farmacokinetische principes van belang. Daarnaast is het essentieel om onderscheid te maken tussen verschillende typen infecties en hierop het individuele antibioticabeleid af te stemmen. Om het antibioticabeleid verder te kunnen optimaliseren is samenwerking tussen tandartsen en artsen van groot belang.

International audience; Gram-positive anaerobic cocci (GPAC) account for 24%\textendash31% of the anaerobic bacteria isolated from human clinical specimens. At present, GPAC are under-represented in the Biotyper MALDI-TOF MS database. Profiles of new species have yet to be added. We present the optimization of the matrix-assisted laser desorption\textendashionization time-of-flight mass spectrometry (MALDI-TOF MS) database for the identification of GPAC. Main spectral profiles (MSPs) were created for 108 clinical GPAC isolates. Identity was confirmed using 16S rRNA gene sequencing. Species identification was considered to be reliable if the sequence similarity with its closest relative was >=98.7%. The optimized database was validated using 140 clinical isolates. The 16S rRNA sequencing identity was compared with the MALDI-TOF MS result. MSPs were added from 17 species that were not yet represented in the MALDI-TOF MS database or were under-represented (fewer than five MSPs). This resulted in an increase from 53.6% (75/140) to 82.1% (115/140) of GPAC isolates that could be identified at the species level using MALDI-TOF MS. An improved log score was obtained for 51.4% (72/140) of the strains. For strains with a sequence similarity \textless98.7% with their closest relative (n~=~5) or with an inconclusive sequence identity (n~=~4), no identification was obtained by MALDI-TOF MS or in the latter case an identity with one of its relatives. For some species the MSP of the type strain was not part of the confined cluster of the corresponding clinical isolates. Also, not all species formed a homogeneous cluster. It emphasizes the necessity of adding sufficient MSPs of human clinical isolates.

The pathogenesis of periodontitis and of rheumatoid arthritis show remarkable similarities. There is a distinct degree of co-existence between the 2 diseases. The prevalence of periodontitis is more pronounced in rheumatoid arthritis patients and the prevalence of rheumatoid arthritis is more pronounced in periodontitis patients. At present, a positive influence of periodontal treatment on the rheumatoid arthritis disease activity or of rheumatoid arthritis drug treatment on periodontitis is not sufficiently supported by clinical research. Periodontitis may play a role in unsatisfactory therapy response in some rheumatoid arthritis patients.

BACKGROUND AND OBJECTIVE: Porphyromonas gingivalis is an oral pathogen strongly associated with destruction of the tooth-supporting tissues in human periodontitis. Gingival fibroblasts (GF) and periodontal ligament fibroblasts (PDLF) are functionally different cell types in the periodontium that can participate in the host immune response in periodontitis. This study aimed to investigate the effects of viable P. gingivalis on the expression of genes associated with inflammation and bone degradation by these fibroblast subsets.MATERIAL AND METHODS: Primary human GF and PDLF from six healthy donors were challenged in vitro with viable P. gingivalis W83 for 6 h. Gene expression of inflammatory cytokines in GF and PDLF was analyzed using real-time PCR, and protein expression was analyzed using ELISA.RESULTS: Viable P. gingivalis induced a strong in vitro inflammatory response in both GF and PDLF. We found increased gene expression of interleukin (IL)-1beta, IL-6, IL-8, tumor necrosis factor-alpha, monocyte chemotactic protein-1 and regulated upon activation, normal T-cell expressed and secreted (RANTES). Macrophage colony-stimulating factor was induced and the expression of osteoprotegerin was decreased in GF, but not in PDLF. In nonchallenged cells, a higher level of expression of IL-6 was observed in GF than in PDLF. Between individual donors there was large heterogeneity in responsiveness to P. gingivalis. Also, in each individual, either GF or PDLF was more responsive to P. gingivalis.CONCLUSION: Considerable heterogeneity in responsiveness to P. gingivalis exists both between GF and PDLF and between individuals, which may be crucial determinants for the susceptibility to develop periodontitis.

Background: Receptors for the Fc part of IgG (Fc gamma RIIa) on polymorphonuclear leukocytes (PMN) mediate phagocytosis and cell activation. Previous results show that one of the genetic variants of the Fc gamma RIIa, the 131 H/H, is associated with more periodontal breakdown than the R/R. This may be due to hyper-reactivity of the H/H-PMNs upon interaction with bacteria.Aim: To study whether the Fc gamma RIIa genotype modifies the PMN reactivity in periodontitis patients.Material and Methods: A cohort of 98 periodontitis patients was genotyped. From these, 10 H/H and 10 R/R consented to participate. PMNs were incubated with immune serum-opsonized Actinobacillus actinomycetemcomitans (A.a.). Phagocytosis, degranulation (CD63 and CD66b expression), respiratory burst and elastase release were assessed.Results: Patients of the H/H genotype showed more bone loss than those with the H/R or R/R genotype (p=0.038). H/H-PMNs phagocytosed more opsonized A.a. than did R/R-PMNs (p=0.019). The H/H-PMNs also expressed more CD63 and CD66b than did the R/R-PMNs (p=0.004 and 0.002, respectively) and released more elastase (p=0.001).Conclusions: The genotyping results confirm previous reports that more periodontal destruction occurs in the H/H genotype than in the H/R or R/R genotype. The functional studies indicate a hyper-reactivity of the H/H-PMN in response to bacteria, which may be one of several pathways leading to more periodontal breakdown.

BACKGROUND: Leukaemic patients receiving intensive chemotherapy and patients undergoing autologous stem-cell transplantation (ASCT) are routinely screened for oral foci of infection to reduce infectious complications that could occur during therapy. In this prospective study we assessed the effect of leaving chronic oral foci of infection untreated on the development of infectious complications in intensively treated haematological patients.METHODS: We included and prospectively evaluated all intensively treated leukaemic patients and patients undergoing ASCT who were referred to our medical centre between September 2012 and May 2014, and who matched the inclusion/exclusion criteria. Acute oral foci of infection were removed before chemotherapy or ASCT, whereas chronic oral foci were left untreated.RESULTS: In total 28 leukaemic and 35 ASCT patients were included. Acute oral foci of infection were found in 2 leukaemic (7%) and 2 ASCT patients (6%), and chronic oral foci of infection in 24 leukaemic (86%) and 22 ASCT patients (63%). Positive blood cultures with microorganisms potentially originating from the oral cavity occurred in 7 patients during treatment, but were uneventful on development of infectious complications.CONCLUSIONS: Our prospective study supports the hypothesis that chronic oral foci of infection can be left untreated as this does not increase infectious complications during intensive chemotherapy.British Journal of Cancer advance online publication, 22 March 2016; doi:10.1038/bjc.2016.60 www.bjcancer.com.

Background: Bacteria play an essential role in the aetiology of periodontitis. Most bacterial species isolated from subgingival plaque are indigenous to the oral cavity. Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis are detected infrequently in periodontal health, which makes these species prime candidates to study person-to-person transmission. The aim of the present study was to review the literature on transmission of these periodontal bacterial species. Method: We review the literature on bacterial typing techniques and summarize the information on clonal distribution of A. actinomycetemcomitans and P. gingivalis in family units based on different typing techniques in order to establish the likelihood for person-to-person transmission of these periodontal pathogens. Results: Vertical transmission of A. actinomycetemcomitans is estimated to be between 30% and 60%, whereas vertical transmission of P. gingivalis has rarely been observed. Horizontal transmission between spouses ranges between 14% and 60% for A. actinomycetemcomitans and between 30% and 75% for P. gingivalis. There is some evidence to show that cohabitation with a periodontitis patient influences the periodontal status of the spouse; however, substantially more information is needed to prove this hypothesis. Conclusions: Transmission of putative periodontal pathogens between family members has been shown. The clinical consequences of these events have been poorly documented. Based on the current knowledge, screening for and prevention of transmission of specific virulent clones of A. actinomycetemcomitans may be feasible and effective in preventing some forms of periodontal disease. P. gingivalis is usually recovered from diseased adult subjects, and transmission of this pathogens seems largely restricted to adult individuals. Horizontal transmission of P. gingivalis may therefore be controlled by periodontal treatment involving elimination or significant suppression of the pathogen in diseased individuals and by a high standard of oral hygiene.

Background and Aim: Antimicrobial resistance of periodontal pathogens towards currently used antibiotics in periodontics has been investigated in a previous study. Microbial resistance in the periodontal microflora was more frequently observed in Spanish patients in comparison with Dutch patients. The aim of the present study was to compare antimicrobial susceptibility profiles of five periodontal bacteria isolated from periodontitis patients in Spain and in the Netherlands. Material and Methods: Subgingival plaque samples from adult patients with periodontitis were collected and cultured on selective and non-selective plates. Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Micromonas micros were isolated and used for minimal inhibitory concentration tests using the Epsilometer (E-test) technique. Eight different antibiotics were tested on all bacterial isolates. MIC50 and MIC90 values for each antibiotic and each species were determined and the percentage of resistant strains was calculated. Results: Significantly higher MIC values were noted in Spanish strains of F. nucleatum for penicillin, ciprofloxacin, of P. intermedia for penicillin, amoxicillin and tetracycline, of M. micros for tetracycline, amoxicillin and azithromycin, and of P. gingivalis for tetracycline and ciprofloxacin. Based on breakpoint concentrations, a higher number of resistant strains in Spain were found in F. nucleatum for penicillin, amoxicillin and metronidazole, in Prevotella intermedia for tetracycline and amoxicillin, and in A. actinomycetemcomitans for amoxicillin and azithromycin. Resistance of P. gingivalis strains was not observed for any of the antibiotics tested both in Spain and the Netherlands. Conclusions: Differences exist in the susceptibility profiles of periodontal pathogens isolated from periodontitis patients in Spain and in the Netherlands. This implicates that antibiotic susceptibility testing is necessary to determine efficacy of antimicrobial agents. Also, clinical studies with antibiotics should take these differences into account. The information from the present study indicates that it may not be possible to develop uniform protocols for usage of antibiotics in the treatment of severe periodontitis in the European Union.

Background: A major aspect of the adaptive host response in periodontitis is the production of antibodies. Several risk and susceptibility factors for periodontitis, including smoking, age and composition of the subgingival microflora, have also been suggested to influence antibody production.Aim: The present study was conducted to investigate plasma levels of immunoglobulin (Ig) G, A and M antibodies in periodontitis patients of Caucasian European heritage in relation to disease severity, smoking, diagnosis and prevalence of periodontopathogens.Methods: In this study, 29 patients with severe periodontitis, 51 with moderate periodontitis and 55 controls without periodontal destruction were enrolled. From the total of 80 patients, 18 were diagnosed with aggressive periodontitis and 62 with chronic periodontitis. Total IgG, IgA and IgM as well as IgG isotypes were analyzed in plasma samples.Results: Levels of total IgG, IgA and IgM were not different between patients and controls; however, in periodontitis, higher levels of IgG1 and IgG2 were observed. Smoking appeared to be significantly and inversely related to antibody levels in periodontitis, in particular for total IgG and IgG2. The absence of an elevated total IgG and IgG2 in smoking patients was irrespective of severity, prevalence of periodontal pathogens and diagnosis. The elevation of total IgG and IgG1 and IgG2 in non-smoker periodontitis patients was observed in patients with moderate periodontitis and even greater in patients with severe periodontitis, but was independent whether patients were infected with Actinobacillus actinomycetemcomitans or Porphyromonas gingivalis and independent of diagnosis. Clinically, it was observed that patients who smoked had more periodontal bone loss; the current findings on antibody levels may be one of several mechanisms related to more extensive periodontal breakdown in smoker patients.Conclusion: The current study shows that non-smoker periodontitis patients have higher levels of total IgG and IgG2 than smoker periodontitis patients.

Aim: This study evaluated the microbial effects of a newly formulated mouthwash (Halita((R)) ) on oral halitosis patients. Methods: Forty subjects were included in this dual-centre, double-blind, placebo-controlled parallel study. Inclusion and exclusion criteria were used to select patients. At baseline and at 2 weeks post-treatment, full-mouth organoleptic odor scores, level of volatile sulphur compounds (VSC) and the Winkel Tongue Coating Index were recorded. Standardized samples of tongue coating, saliva and subgingival plaque were microbiologically investigated. Participants were randomly assigned to the test or placebo groups. Results: High prevalences were observed for Fusobacterium nucleatum , Prevotella intermedia and Porphyromonas gingivalis in tongue coating, saliva and subgingival plaque samples. A significant positive correlation between baseline total counts of P. gingivalis in saliva samples and organoleptic and VSC scores was found. Two weeks post-treatment there was a reduction in total anaerobic counts in all samples in the test group. A significant positive correlation was observed between the reduction in total counts in saliva samples and the reduction in organoleptic scores in the test group. Significant reductions in total counts and proportions of F. nucleatum and total counts of P. intermedia in tongue coating samples were observed in the test group. Conclusions: The test mouthwash demonstrated efficacy in reducing the microbiological parameters in three oral niches in moderate to severe halitosis patients without periodontitis, and this was correlated with the improvements in organoleptic and VSC scores but not with the tongue coating scores.

Objectives: The aim of this double-blind, parallel study was to test the clinical efficacy of a newly developed mouthrinse in the treatment of oral halitosis in patients without periodontitis. Material and methods: Forty volunteers, recruited in two centers, participated in this study. Patients were selected on the basis of (1) halitosis of oral origin, (2) full-mouth organoleptic score >1, using an arbitrary 0-5 scale, (3) level of volatile sulfur compounds (VSC) >170 parts per billion (ppb) and (4) Winkel tongue coating index (WTCI) > 4 (0-12). Intervention included gargling with a mouthrinse containing chlorhexidine (0.05%), cetylpyridinium chloride (0.05%) and zinc-lactate (0.14%) or with a placebo mouthrinse without active ingredients. At days 0 and 14 clinical variables were assessed in order of performance: (1) organoleptic assessments, (2) levels of VSC, and (3) WTCI. Results: Treatment with the active mouthrinse resulted in a significant mean reduction in the organoleptic score from 2.8 to 1.5 (p

Background: The effect of smoking on the prevalence of periodontal pathogens after periodontal treatment is still not clear. Some studies found no effect of the smoking status on the prevalence of periodontal pathogens after therapy, whereas others did. The aim of this retrospective study was to investigate the influence of smoking on the treatment of periodontitis and the composition of the subgingival microflora. Method: The study included 59 periodontitis patients (mean age 41.5 years): 30 smokers and 29 nonsmokers. The treatment consisted of initial periodontal therapy and, if necessary, surgery and/or antibiotics. Clinical and microbiological data were obtained before and after treatment at the deepest site in each quadrant. A pooled sample was analysed for the presence of Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotalla intermedia (Pi), Bacteroides forsythus (Bf), Fusobacterium nucleatum (Fn) and Peptostreptococcus micros (Pm). Results: For smokers and nonsmokers a significant improvement of the clinical condition was found after treatment. A decrease could be assessed for bleeding on probing (smokers: 0.46; nonsmokers: 0.52) and probing pocket depth (PPD) (smokers: 1.64 mm; nonsmokers: 2.09 mm). Furthermore, both groups showed gain of attachment (smokers: 0.68 mm; nonsmokers: 1.46 mm). No significant difference in bleeding on probing and PPD reduction was found between smokers and nonsmokers. In contrast, nonsmokers showed significantly more gain of attachment than smokers. The microbiological results revealed no differences in the prevalence of the various bacteria between smokers and nonsmokers before treatment. After treatment in nonsmokers, a significant decrease was found in the prevalence of Aa (11–3), Pg (17–7), Pi (27–11), Bf (27–11), Fn (28–20) and Pm (27–17). In smokers, a significant decrease could be shown only for the prevalence of Pg (15–5). Conclusions: Nonsmokers showed more gain of attachment and a greater decrease in the prevalence of periodontal bacteria as compared to smokers. The phenomenon that among smokers, more patients remain culture positive for periodontal pathogens after therapy, may contribute to the often observed unfavourable treatment results in smoker periodontitis patients.

Objectives: Current literature is ambivalent on the use of barrier membranes for regeneration of intraosseous defects. One of the reasons for unpredictable results may be related to infection before, during and after the surgical procedure. Therefore, the purpose of the present controlled study was to evaluate both the use of membranes (MEM) and antibiotics (AB), separately and in combination. Methods: In all, 25 patients with two intraosseous periodontal defects each were randomized in two groups: AB+ group receiving systemic antibiotics (n=13) and AB-group without antibiotics (n=12). After raising flaps and after debridement, both defects in each patient were covered by a bioresorbable membrane (MEM+). However, just before suturing the flaps in a coronal position, the membrane over one of the two defects was removed at random (MEM-). This protocol resulted in four groups of defects: (i) MEM- AB-; (ii) MEM+ AB-; (iii) MEM- AB+; (iv) MEM+ AB+. Patients were monitored clinically and microbiologically for 1 year. Data were analyzed in repeated measures ANCOVA's and adjusted means for clinical variables were obtained from the final statistical model. Results: Reduction in probing pocket depth (PPD) at 12 months postoperatively varied between 2.54 and 3.06 mm between the four treatment modalities, but overall no main effect of MEM or AB was found. Gains in probing attachment level (PAL) at 12 months postoperatively varied between 0.56 and 1.96 mm for the 4 treatments. In the overall analysis for PAL, no main effect of MEM or AB was found. Gains in probing bone level (PBL) 12 months postoperatively ranged from 1.39 to 2.09 mm between the treatment groups. Again, overall, no main effects of MEM or AB were found for PBL. Explorative statistical analyses indicated that smoking and not MEM or AB is a determining factor for gain in PBL (P=0.0009). Nonsmokers were estimated to gain 2.04 mm PBL compared to 0.52 mm in smokers. The prevalence of several periodontal pathogens, at the day of surgery or postoperatively, and specific defect characteristics, were not determining factors for gain in PAL and PBL. Conclusions: Neither the application of barrier membranes nor the use of systemic antibiotics showed an additional effect over control on both soft and hard tissue measurements in the treatment of intraosseous defects. In contrast, smoking was a determining factor severely limiting gain in PBL in surgical procedures aimed at regeneration of intraosseous defects.

Background: Periodontal disease in Down's syndrome (DS) is generally characterized by a high degree of bone loss. Bone loss of 5 mm or more is observed in 70% of these subjects. Among DS subjects, considerable differences in disease progression occur. So far, no studies have been conducted in which specific properties of the subgingival microflora have been related to the condition observed. Aims: To investigate (1) the subgingival microflora in DS subjects and other mentally retarded (control) individuals which were matched to the utmost and (2) to investigate the subgingival microflora of a "low-risk" and a "high-risk" group formed in DS subjects. Material and Methods: 17 DS subjects and 17 control subjects were matched with respect to age, plaque level and bleeding on probing. In addition, the DS group was divided in a "low-risk" group (0-2 teeth lost due to periodontal disease n = 6) and a "high-risk" group (6-13 teeth lost due to periodontal disease n = 11). Prevalence and proportions of the putative periodontal pathogens Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus, Peptostreptococcus micros, Fusobacterium nucleation and Campylobacter rectus in the subgingival plaque were determined using anaerobic culture techniques. No differences in the prevalence of distinct suspected periodontopathic bacteria and bacterial subgingival composition between the DS group and the control group could be established. Also no differences in the prevalence of the seven investigated microbial species between the "low-risk" and the "high-risk" group were observed. Conclusions: Because of the lack of differences in microflora between the DS group and the control group, a specific effect of the microbiological composition in the periodontal status of subjects with DS can be excluded in this population. Host factors constitute the more likely explanation of the differences observed in DS.

Background: The etiology of root caries is thought to be identical to coronal caries, though root caries seem to be more complicated because of the higher susceptibility of exposed roots (dentin) by periodontal therapy to demineralization than intact enamel. This implies that mutans streptococci are the most likely pathogens in the development of root caries. Although it is known that both the numbers of mutans streptococci and the frequency of isolation in root caries lesions are negatively correlated with the distance from the gingival margin, the subgingival sulcus has not been considered a possible habitat for mutans streptococci. However, subgingival occurence of mutans streptococci in both untreated and treated periodontal patients has not been documented well in the literature. Objective: To investigate the presence and levels of mutans streptococci in the subgingival plaque of patients (n=154) in different stages of periodontal therapy. The subgingival sulcus may be a possible habitat for mutans streptococci. This localisation of mutans streptococci may be of importance in the development of root caries after periodontal surgery. Materials and methods: In this cross-sectional study, subgingival plaque samples from 154 consecutive adult periodontitis patients were tested for presence and levels of mutans streptococci and putative periodontal pathogens by anaerobic cultures. These patients were divided into 3 groups based on their stage of periodontal treatment: (1) untreated patients; (2) patients after initial periodontal therapy only; (3) patients in the maintenance phase who not underwent periodontal surgery; (4) patients after periodontal surgery. Results: The prevalence of mutans streptococci in the 4 study groups varied from 82% in untreated patients to 94% in maintenance patients. The mean proportion of mutans streptococci was 6.65% in maintenance patients versus 1.86% in untreated patients (p=0.005) and 2.51% in patients after scaling and root planing (p=0.041). Conclusions: The subgingival area is a microbial habitat for mutans streptococci that may be of importance in the development of root caries in periodontitis patients.

The purpose of this study was to compare the subgingival microbiota of two geographically distinct patient populations using identical clinical and bacteriological methods. Adult patients with a diagnosis of periodontitis were consecutively selected according to pre-defined clinical criteria. Microbiological samples were taken from the deepest four sites with bleeding. The samples were plated on blood agar plates, for the determination of the total anaerobic counts and identification of specific bacterial pathogens, and on TSBV and McConkey for isolation of Actinobacillus actinomycetemcomitans and enteric rods, respectively. Thirty-one patients in Spain and 30 patients in The Netherlands were selected. Both patient groups showed similar clinical characteristics, both in terms of age, gender and periodontal clinical variables. A. actinomycetemcomitans was significantly more prevalent (23.3% vs. 3.2%) in the Dutch group, while Porphyromonas gingivalis was significantly more prevalent (64.5% vs. 36.7%) in the Spanish group. Bacteroides forsythus and most commensal periodontal pathogens showed similar prevalences, except Peptostreptococcus micros that was significantly more frequent in the Dutch group (96.7% vs. 74.2%). In summary, the subgingival microbiota from the Spanish group was characterised by a high prevalence of P. gingivalis and low of A. actinomycetemcomitans, while the flora from the Dutch group was characterised by a high prevalence of A. actinomycetemcomitans and P. micros.

Background: We describe a simple, non-invasive mouthwash sampling method for rapid DNA isolation to detect cytokine gene polymorphisms. In the present paper, interleukin-1 beta (IL-1B) and interleukin-1 receptor antagonist (IL-1RN) gene polymorphisms were studied. Methods: Two mouthwash samples and blood samples were collected from 11 healthy individuals. The second mouthwash sample was stored for 7 days at room temperature. Polymerase chain reaction amplification was used to identify a bi-allelic polymorphism at position +3953 in the IL-1B gene and a variable number of tandem repeats (VNTR) polymorphism in the IL-1RN gene. Results: Our results show that the typing of these cytokine gene polymorphisms using DNA isolated from mouthwash samples did not differ from those obtained by a phenol/chloroform isolation method from EDTA anti-coagulated blood. Moreover, reliable results from mouthwash samples were obtained after storage for at least 7 days at room temperature. Conclusions: Mouthwash can be the method of choice to study gene polymorphisms in periodontitis and other chronic inflammatory diseases.

Background: Peri-implantitis is a risk factor for implant loss. Late bacterial infection of the peri-implant tissues and loss of alveolar bone in edentulous patients is caused by commensal oral anaerobic bacteria. In partially edentulous patients, Porphyromonas gingivalis and occasionally Actinobacillus actinomycetemcomitans are associated with peri-implantitis lesions. Aims: To investigate the microbiology of a peri-implantitis case in an edentulous patient. Methods: Anaerobic culture techniques and selective culture techniques for A. actinomycetemcomitans were used to study the peri-implant microflora at sites with and without bone loss. Results: An anaerobic peri-implant microflora with several putative periodontal pathogens was found at sites with bone loss. Furthermore, a metronidazole-resistant A. actinomycetemcomitans was isolated. The A. actinomycetemcomitans infection did not respond to systemic doxycycline therapy, despite good susceptibility in vitro. Conclusions: The present case of severe A. actinomycetemcomitans-associated peri-implantitis shows the importance of pre-operative infection control. The findings in this case show that remaining teeth affected by periodontitis can be a serious risk factor for peri-implantitis.

Background, aims: Congenital neutropenia is one condition that may predispose for destructive periodontal disease at a young age. In this report, we describe the microbiology of 3 adolescent patients with congenital neutropenia two of whom suffered from severe periodontitis. Method: Microbiological testing of the parents was also performed in 1 case. DNA fingerprinting was used to study transmission of putative periodontal pathogens in this case. From 1 patient with periodontitis, Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis were isolated; a 2nd periodontitis patient was infected with P. gingivalis. A 3rd patient had gingivitis only and no A. actinomycetemcomitans or P. gingivalis were found. Results: Using the amplified fragment length polymorphism DNA fingerprinting technique, bacterial transmission between the father and a patient was shown for A. actinomycetemcomitans but not for P. gingivalis.

Concentrations and output of lactoferrin and of low-Mr mucin MG2 were determined in saliva of subjects suffering from Actinobacillus actinomycetemcomitans-associated periodontal disease and healthy subjects. Periodontal patients were clinically examined and a microbiological sample was taken from the deepest bleeding pockets in each quadrant. The number of viable A. actinomycetemcomitans was determined in the sampled sites of each patient. The MG2 output in the diseased subjects (13.6 microg protein/min) was decreased at least by a factor three compared to periodontal healthy subjects (44.3 microg protein/min). On the other hand, output of lactoferrin was not significantly different in healthy (9.5 microg/min) and diseased subjects (7.6 microg/min). Western analyses demonstrated a higher iron-saturation of lactoferrin in diseased subjects in comparison with the healthy subjects. Lactoferrin degrading enzymes, probably derived from microbial sources, could be detected in saliva of the periodontally diseased subjects, but not in saliva of healthy subjects. The combination of iron-saturation and degradation of lactoferrin suggests that anti-microbial properties of lactoferrin are diminished in periodontitis patients. Moreover, the low concentration of mucin MG2 suggests a decline in mucin defence and consequently a higher susceptibility for oral infection. A negative correlation (r= -0.4, p0.05) between the number of subgingival A. actinomycetemcomitans and lactoferrin in saliva suggested that low concentrations of lactoferrin favour the growth of the bacterium. These data indicate that a decline in the salivary defence system might increase the risk for oral infection by A. actinomycetemcomitans.

Capsular structures of Porphyromonas gingivalis have been correlated to the pathogenicity in animal models. Six polysaccharide capsular serotypes have recently been described in P. gingivalis. In the present study, virulence of the P. gingivalis strains of the six capsular serotypes was compared with strains of the non-capsular serotype of P. gingivalis in a mouse model. All 18 encapsulated strains caused a spreading type of infection with exudate, often accompanied by ulceration and necrosis of the skin and a significant loss of body weight. All three non-encapsulated strains tested induced localized abscesses, although a spreading type of infection was occasionally observed. Illness of the mice was almost always associated with recovery of encapsulated P. gingivalis from blood, kidneys, lungs and spleens. A statistically significant contrast was found in the average weight change during the experiment between the mice infected with the K- serotype strains and the mice infected with the encapsulated P. gingivalis strains. Differences in severity of illness and the recovery of P. gingivalis from mouse tissues were recorded between strains of the same serotype. The differences in virulence within a capsular serotype suggest that the capsule is an important but not the only determining virulence factor for P. gingivalis.

Previous reports have described six serotypes based on K antigens in Porphyromonas gingivalis strains. The purpose of the present study was to investigate the prevalence and distribution of these serotypes in 185 patients with P. gingivalis-associated periodontitis. Polyclonal rabbit antisera, raised against each of the different type strains, were used in double-immunodiffusion and immunoelectrophoresis assays. In addition, a subset of 76 strains was investigated for the presence of capsular structures by means of the India ink and Bruce White staining techniques. These strains were also tested for auto-aggregation in phosphate-buffered saline (PBS). All six K serotypes were present in the study sample. In total, 84 (45.4%) patients were colonized with a K-typeable P. gingivalis strain with a predominance of types K5 (12%) and K6 (23.2%). A correlation was found between arbitrary age categories and the prevalence of currently known K serotypes, which were found in 60% of patients aged 12 to 30 years, in 49% of patients aged 31 to 50, and in 25% of patients aged 51 to 70 years. In the subset of 76 P. gingivalis strains, 32 (42.1%) were K-typeable. Fifty-three strains (69.7%) showed microscopic evidence of encapsulation, suggesting the existence of K serotypes other than K1 to K6. Twenty-one strains (27.6%) auto-aggregated in PBS and were not K-typeable, nor did they show any evidence of encapsulation. It was concluded that the majority of clinical P. gingivalis isolates is encapsulated and that encapsulation is associated with the presence of a K antigen. Auto-aggregation seems to be associated with the absence of a capsular structure and, consequently, the absence of a K antigen.

Recently van Winkelhoff et al. (1) described 3 novel serotypes in virulent Porphyromonas gingivalis strains, which were based on different polysaccharide antigens. These antigens probably represent capsular structures and have been designated K1, K2 and K3. In the present study we report on 3 novel capsular serotypes, which are represented by P. gingivalis strains ATCC 49417, HG 1690 and HG 1691. The strains, isolated from patients with periodontitis, showed obvious encapsulation in wet India ink preparations. Thermostable antigens could be detected in the supernatant fractions of autoclaved cells. These antigens appeared to be negatively charged, sensitive to periodate degradation, and resistant to proteinase K treatment. On the basis of these characteristics we conclude that the antigens are probably extra-cellular polysaccharides representing a bacterial capsular structure. These K-antigens did not cross-react with K1, K2 or K3 immune-sera of P. gingivalis, with the exception of the K2 antiserum, which partially recognized K5- and K6-antigens. In contrast, K5 and K6 antisera did not react with the K2-antigen. After absorbtion of the K2 antiserum with cells of strains HG 1690 (K5) and HG 1691 (K6) cross-reactivity was no longer present. We propose these novel serotypes to be designated: K4 (ATCC 49417), K5 (HG 1960) and K6 (HG 1691).

Background Early diagnosis and treatment of rheumatoid arthritis (RA) is of paramount importance for achieving a better disease outcome. Seropositive arthralgia patients (SAP), positive for anti citrullinated protein antibody (ACPA) and/or rheumatoid factor (RF) with (a history of) arthralgia, have a “high risk” of developing RA. Therefore it is necessary to understand which preclinical immunological factors play a role in the progression towards RA in SAP since this could help identify individuals earlier and result in early intervention. Objectives This prospective study aimed at identifying whether altered regulatory T-cells (Tregs) levels and subsets, besides a broadened ACPA response, are biomarkers for RA development in SAP. Methods Thirty-four consecutive SAP were prospectively assessed every 6 months for at least 2 years. Every visit, peripheral blood mononuclear cells (PBMCs) were isolated and stored. At inclusion, peripheral Treg (CD4 + CD25 + FoxP3 + ) levels and Treg subsets, defined as CD45RA + FoxP3 low naive Treg cells (Fr I), CD45RA – FoxP3 high activated Treg cells (Fr II) and cytokine producing CD45RA – FoxP3 low non-Treg cells (Fr III), were compared to age and sex matched healthy controls (HC, n=16) and treatment naive RA patients (n=12). SAP that developed RA during follow-up were compared to non-switchers on Treg levels and functional Treg subsets. To assess broadening of the IgG and IgA ACPA repertoire in serum, reactivity was measured against citrullinated peptides from fibrinogen, α-enolase and vimentin. Results Total Treg levels were similar between HC, SAP and RA patients. Although functional Treg subsets Fr I and Fr II were comparable between groups, Fr III was increased in SAP compared to HC (p=0.01). Fourteen (41%) SAP developed RA during follow-up. Their Treg levels were comparable to those of non-switched SAP. At RA diagnosis, Treg levels in switched SAP were not changed compared to 6 months before. Switched SAP displayed an extended IgG ACPA repertoire compared to non-switched SAP (p=0.046). Conclusions Total and functional Treg levels do not indicate development of RA in SAP, in contrast to an extended IgG and IgA ACPA repertoire. Hence, there is presumably no contribution of altered Treg levels leading to the loss in suppression of autoimmunity in RA pathology. Disclosure of Interest None declared

Actinobacillus actinomycetemcomitans is a facultative anaerobic bacterium that displays moderate susceptibility to metronidazole and this study was undertaken to identify the factors involved. A. actinomycetemcomitans appeared two to four times less susceptible to metronidazole when grown in air supplemented with 5% CO 2 than under anaerobic conditions. Ferredoxin-linked pyruvate: oxidoreductase activity was absent but each strain exhibited nitroreductase activity which corresponded directly with uptake of metronidazole and susceptibility to the drug under anaerobic conditions but not in air supplmented with 5% CO 2 . Nitroreductase activity therefore appears responsible for the susceptibility of A. actinomycetemcomitans to metronidazole

Actinobacillus actinomycetemcomitans is an important pathogen in the etiology of severe periodontitis. For epidemiological studies on the prevalence of certain pathogenic clones and transmission of this bacterium, adequate typing methods are necessary. The purpose of this study was to compare six different typing methods for A. actinomycetemcomitans. Five reference strains and 27 fresh clinical isolates from periodontitis patients were used. Serotyping showed 12 serotype a strains, 13 type b strains, 6 type c strains, and 1 nontypeable strain. Biotyping on the basis of the fermentation of mannose, mannitol, and xylose resulted in six biotypes. Antibiogram typing was evaluated by measuring the inhibition zones of seven antibiotics in agar diffusion tests. With this method eight main types which could be further differentiated into 15 subtypes were found. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of outer membrane proteins were similar among all isolates tested. Restriction endonuclease analysis (REA) of whole chromosomal DNA resulted in five main types. These five main types were further differentiated into 24 subtypes on the basis of DNA fragment differences in the high-molecular-weight region. Hybridization of DNA fragments with ribosomal DNA (ribotyping) resulted in 22 to 24 different types, depending on the restriction endonuclease used. Ribotype patterns were easy to interpret and provided an univocal distinction between different strains compared with REA results. When applied to epidemiologically related isolates, all methods were able to discriminate two clonal types among five isolates from five children from one family. We conclude that serotyping, biotyping, and outer membrane patterns were reproducible but had a low discriminatory potential. REA and ribotyping were reproducible and gave the highest number of distinct types. When the DNA typing methodis were compared, all strains tested could be distinguished. These findings confirm the heterogeneity found within the species A. actinomycetemcomitans.

In this study we determined the prevalence and distribution of Porphyromonas endodontalis in 26 families consisting of 107 subjects. P. endodontalis was present in 24% of the investigated subjects and was recovered most often from the dorsum of the tongue (50%). Isolation was also possible from the tonsils, the buccal mucosa, the saliva and the periodontal pocket. The usefulness of restriction endonuclease analysis as a typing method for this particular species was investigated by typing 19 isolates from unrelated individuals. All these isolates had unique restriction endonuclease patterns. The observed heterogeneity indicates that restriction endonuclease analysis is a sensitive measure of genetic dissimilarity between P. endodontalis isolates and is able to characterize individual isolates. Application of restriction endonuclease analysis to the obtained clinical isolates in this study shows the possibility of the presence of multiple clonal types within one subject. The DNA patterns of all P. endodontalis isolates from unrelated individuals were found to be distinct. In 3 families the DNA patterns of isolates from the mother and her child were indistinguishable. These data indicate the possibility of intrafamilial transmission of P. endodontalis.

Until recently, black-pigmented Gram-negative anaerobes were classified as 'black-pigmented Bacteroides'. At present, 11 distinct species are recognized in this group. Because of major differences with Bacteroides fragilis, the type species of the genus Bacteroides, new genera have been proposed: Porphyromonas for three asaccharolytic species, and Prevotella for the saccharolytic species. Typing methods have been developed for some species of black-pigmented Gram-negative anaerobes. These include biotyping and serotyping, but relatively few types can be distinguished with these methods. Recently, DNA restriction endonuclease analysis has been used for typing of P. gingivalis, Pr. intermedia and P. endodontalis strains. Great heterogeneity was observed within all three species. This typing method can be useful for epidemiological studies.

The in vitro susceptibilities of Actinobacillus actinomycetemcomitans to 14 antimicrobial combinations were studied by using the checkerboard titration technique. The results, expressed as the range of the fractional inhibitory concentration indices, were as follows: for metronidazole or its hydroxymetabolite combined with cefixime, 0.2 to 0.6; for moxalactam, 0.2 to 0.6; for penicillin G, 0.3 to 0.6; for tobramycin, 0.8 to 2.0; for erythromycin, 0.8 to 1.7; for ciprofloxacin, 0.2 to 0.6; for tetracycline, 0.8 to 1.2. Our observations indicated that the beta-lactam antibiotics as well as ciprofloxacin act synergistically with both metronidazole and its hydroxymetabolite against A. actinomycetemcomitans. Synergistic interactions were independent of the individual MICs of the antibiotics tested. Erythromycin, tobramycin, and tetracycline combined with either metronidazole or its hydroxymetabolite showed additive to indifferent effects against the five strains of A. actinomycetemcomitans, with the fractional inhibitory concentration indices ranging from 0.8 to 2.0. A. actinomycetemcomitans was found to be highly susceptible to ciprofloxacin (MIC of ciprofloxacin for 90% of strains tested, 0.010 micrograms/ml) and cefixime (MIC of cefixime for 90% of strains tested, 0.8 micrograms/ml). The results indicate that in patients who are allergic to penicillin, cefixime and ciprofloxacin may be useful alternative antibiotics in combination with metronidazole for the treatment of A. actinomycetemcomitans-associated periodontitis.

Parodontitis is een multifactoriele ziekte, dat wil zeggen dat verschillende risico’s zijn vastgesteld voor deze aandoening (Page e.a., 1997). Een risicofactor voor parodontitis wordt gedefinieerd als een factor (kan zowel een omgevings- als gedrags- of biologische factor zijn) die aanwezig is voordat de ziekte zich voordoet en waarvan de relatie tot de ziekte bevestigd is in longitudinale studies. De aanwezigheid van een dergelijke risicofactor verhoogt dus de kans op het ontstaan van de ziekte, terwijl de afwezigheid van de risicofactor de kans juist verkleint (Norderyd, 1998).

De geschiedenis van de orale microbiologie begint bij Antoni van Leeuwenhoek in 1683. Hij beschreef met behulp van een eenvoudige microscoop bacterien in tandplaque. Tevens legde hij het verband tussen het aantal bacterien en geringe mondverzorging.

Interactions between metronidazole and amoxicillin, metronidazole and its hydroxymetabolite, and amoxicillin and the hydroxymetabolite of metronidazole were investigated with checkerboard titrations in combination with accurately determined MICs and MBCs. Actinobacillus actinomycetemcomitans was used as the test organism. Synergism was found for all three combinations. Fractional inhibitory concentration indices and fractional bactericidal concentration indices varied from 0.3 to 0.7. These synergistic interactions between these antibiotics may explain the efficacy of the combination of metronidazole and amoxicillin in various bacterial infections, including periodontal disease.

Introduction: In the past, theories on the transmission of Porphyromonas gingivalis between individuals have been based on, among other techniques, restriction enzyme analysis (REA) of bacterial DNA. Currently, amplified fragment length polymorphism (AFLP) may be a more sophisticated alternative. The possibility of automatic pattern analysis and digital storage of the typing data enables the comparison of patterns from a large number of strains in a broad time frame. The aim of this study was to compare REA profiles with AFLP patterns of P. gingivalis strains isolated from periodontitis patients and their spouses. Methods: Forty-two P. gingivalis strains were isolated from different sites in the mouth from six adult patients with periodontitis and their spouses. DNA of the bacterial isolates was subjected to REA and AFLP analysis. Results: One single type of P. gingivalis was found in each individual with both methods, regardless of the site of isolation. Indistinguishable types were found in four of the six couples with both techniques. Different types were found in two couples with both the REA and the AFLP method. Conclusions: The AFLP typing technique confirms earlier observations on the transmission of P. gingivalis between spouses. This new technique can replace REA typing.

Aim: To study transmission of Porphyromonas gingivalis in a population living in a remote area in Southern Java, Indonesia.Material and Methods: Subgingival plaque samples from 167 subjects with varying degrees of periodontal breakdown were obtained and cultured for the presence of P. gingivalis. After extraction and purification of bacterial DNA, amplified fragment length polymorphism technique was applied to genotype the bacterial isolates. Computer-assisted analysis of the bacterial DNA profiles was used to study distribution of P. gingivalis genotypes within family units.Results: One hundred and five of the 167 (63%) subjects were culture positive for P. gingivalis. In total, 371 P. gingivalis isolates were obtained from the 105 subjects. Of the 105 subjects, 30 were siblings representing 13 families. In six of the 13 families (46%), identical P. gingivalis genotypes were found among siblings. In the study group of 105 subjects, 13 married couples were identified of which both spouses were culture positive for P. gingivalis. None of the 13 couples shared an identical P. gingivalis genotype. Twenty P. gingivalis-positive subjects had spouses that were culture negative for P. gingivalis.Conclusions: In this study population, vertical transmission of P. gingivalis has occurred within family units, most likely from parents to children. Transmission of P. gingivalis between spouses could not be established.

Objective: To investigate (1) reduction in the number of microorganisms obtained directly after subgingival instrumentation, (2) rate of bacterial re-colonization during 2 weeks, under supragingival plaque-free conditions. Materials and Method: Effects of subgingival instrumentation were measured at one deep pocket in 22 patients (11 smokers and 11 non-smokers). Immediately after initial therapy, experimental sites, under strict plaque control, were instrumented subgingivally. Microbiological evaluation was performed at pre-instrumentation, immediate post-instrumentation and 1 and 2 weeks post-instrumentation. Results: Mean total anaerobic clolony forming units (CFUs) dropped from 3.9 x 10(6) before to 0.09 x 10(6) immediately following instrumentation. Significant reductions were found for Tannerella forsythia, Micromonas micros, Fusobacterium nucleatum and spirochetes. Significant reductions were not observed for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia and Campylobacter rectus. Except for spirochetes, no reduction in prevalence of specific periodontal bacteria was found immediately after instrumentation. During follow-up, mean total CFU tended to increase. Prevalence of periodontal bacteria further reduced. No effect of smoking was found. Conclusion: Results indicate that subgingival mechanical cleaning in itself, has a limited effect, in actually removing bacteria. The subsequent reduction in prevalence of specific periodontal bacteria shows that it is apparently difficult for these species to survive in treated pockets.

van Winkelhoff AJ. Antibiotics in periodontics: are we getting somewhere? J Clin Periodontol 2005; 32: 1094-1095. doi: 10.1111/j.1600-051X.2005.00830.x. (c) Blackwell Munksgaard 2005.

Objective: The aim of this study was to determine the microbial atmospheric contamination during initial periodontal treatment using a piezoelectric ultrasonic scaler in combination with either high-volume evacuation (HVE) or conventional dental suction (CDS).Methods: The study included 17 treatment sessions, consisting of a 40-min episode of continuous plaque and calculus removal using an ultrasonic unit (EMS). The treatment sessions were carried out in six patients with generalized adult periodontitis and ranged from two to four sessions per patient according to their needs. The use of HVE and CDS was randomly assigned over the sessions within each patient. Before each treatment, the operating room was not used for 15 h. To measure baseline microbial air pollution two Petri dishes containing blood agar were exposed for 10 min to the air. At the start of each treatment session, two Petri dishes were exposed for 5 min at a distance of 40 cm from the mouth of the patients. After 20 min, this procedure was repeated. At a distance of 150 cm, two Petri dishes were exposed for 20 min followed by exposure of two new Petri dishes for the rest of the session. The plates were cultured aerobically and anaerobically for 3 and 7 days, respectively.Results: The mean colony forming units (CFU) before treatment never exceeded 0.6 colonies per plate. At 40 cm, the mean CFU, when considering a period of 40 min, was 8.0 for HVE and 17.0 for CDS. The mean CFU at 150 cm during this period was 8.1 with HVE and 10.3 with the CDS. With reference to the Air Microbial Index the operatory atmosphere was considered to be in a good condition during 40 min of continuous use of the ultrasonic scaler in combination with both HVE and CDS.Conclusion: Within the restrictions of this study, only limited atmospheric microbial contamination is produced when using a piezoelectric ultrasonic scaler.

Synergy between metronidazole and its hydroxymetabolite and between each compound and amoxicillin or tetracycline-HCl was determined against Helicobacter pylori. Metronidazole plus its hydroxymetabolite and either compound combined with amoxicillin showed synergism against all 10 strains of H. pylori tested. Metronidazole plus tetracycline-HCl or the hydroxymetabolite plus tetracycline-HCl acted synergistically against seven and six strains, respectively, acted additively against three strains, and had no additional effect against one strain. These results may help to explain the in vivo efficacies of metronidazole combinations in the treatment of H. pylori-associated gastritis.

Background and description of case: The etiology and pathogenesis of generalized cervical root resorptions is not well understood. In the present report, a case of severe cervical root resorption involving 24 anterior and posterior teeth is presented. The lesions developed within a period of 2 years after the patient had changed to an acid-enriched diet. They extended far into the coronal dentin and were associated with gingival inflammation and crestal bone resorption. However, no generalized clinical attachment loss had occurred. Culturing of subgingival plaque revealed the presence of several putative periodontal pathogens among which Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. Treatment consisted of mechanical debridement supported by systemic antibiotics (amoxycillin plus metronidazole) and dietary advice, Results: Within 1 year after the onset of treatment, all resorptive lesions had repaired by ingrowth of a radio-opaque mineralized tissue. The crestal areas showed radiological evidence of bone repair. 3 years after the onset of therapy, one premolar was extracted and examined histologically. ft appeared that irregularly-shaped masses of woven bone-like tissue had invaded into the domain of the resorbed coronal dentin and were bordered by thin layers of acellular cementum. Conclusion: It is concluded that, in this patient, the cervical resorptions were likely the result of an osteoclastic response extending into the roots because the root-protective role of the junctional epithelium did not develop. We hypothesize that this was due to the combined effects of a periodontopathogenic microflora and a dietary confounding factor.

Background, aims: In order to investigate the role of various putative clinical and microbiological risk markers, a longitudinal study was initiated in a young population deprived of regular dental care. In 1987 all inhabitants in the age range 15-25 years living in a village with approximately 2000 inhabitants at a tea estate on Western Java, Indonesia, were examined clinically and microbiologically. In total, 167 subjects of the original group of 255 adolescents were re-examined in 1994. The material presented in this paper describes the clinical periodontal condition at baseline (1987) and at follow-up (1994), 7 years later. Furthermore, the relationship between progression of the disease and baseline clinical and microbiological data was assessed. Methods: Plaque index (PI), bleeding on probing (BOP), pocket depth (PD), and attachment loss (AL) were scored at the approximal surfaces of the vestibular aspects of all teeth. The number of approximal surfaces of the Ramfjord teeth showing subgingival calculus was recorded. At baseline, the dorsum of the tongue, the buccal gingiva in the upper jaw, the saliva and the deepest bleeding pocket without clinical loss of attachment were sampled for microbiological examination with phase contrast microscopy and indirect immunofluorescence. Results: Mean values at baseline and at follow-up were PI: 1.01 and 1.15, BOP: 0.80 and 1.16, PD 3.26 mm and 3.32 mm, AL: 0.33 mm and 0.73 mm, respectively. All parameters except PD showed a statistically significant increase over the 7-year period. The prevalence of the studied bacteria irrespective of the sample site was: A. actinomycetemcomitans 53%, P. gingivalis 88%, P. intermedia 100%, spirochetes 89% and motile micro-organisms 100%. At the full mouth level, logistic regression showed significant odds ratios for progressive disease with age (1.15), subgingival calculus (1.20) and subgingival presence of A. actinomycetemcomitans (4.61). Presence of any of the selected micro-organisms on the mucous membranes was not related with progressive disease. In order to study local factors to explain local disease activity, each subject was characterized using the sampled pocket, which was the deepest bleeding pocket without LA at baseline, as a single response site per patient. In this constrained design, the main statistical factors associated with progressive disease were presence of motile micro-organisms and the plaque score. Conclusions: This study identified 3 main risk markers for disease progression at the full mouth level: age, amount of subgingival calculus and subgingival presence of A. actinomycetemcomitans.

The aims of this study were to evaluate the clinical and microbiological effects of initial periodontal therapy (IT) and to determine the additional effects of systemic amoxicillin (Flemoxin Solutab) 375 mg TID plus metronidazole 250 mg TID therapy, in patients with adult Actinobacillus actinomycetemcomitans (Aa)-associated periodontitis in conjunction with either Porphyromonas gingivalis (Pg), Bacteroides forsythus (Bf) and/or Prevotella intermedia (Pi). In addition the adverse effects of the antimicrobial therapy were also documented. A total of 22 patients were enrolled. The deepest, bleeding pocket in each quadrant was selected and at these 4 experimental sites clinical measurements and microbiological testing was carried out at baseline, after (IT), i.e., 21 weeks after baseline, and after antimicrobial therapy (AM), i.e., 35 weeks after baseline. At baseline, the mean plaque index (PI) amounted 0.5, 0.1 after IT and 0.3 after systemic AM. The mean bleeding index decreased from 1.6 to 1.2 after IT and a further decrease to 0.7 after AM was noted. Suppuration was completely eliminated after AM. The mean change of probing pocket depth (PPD) after IT amounted 1.4 mm and was further reduced with an additional mean change of 1.1 mm after medication. Clinical attachment gain was 1.1 mm after IT and an additional 0.9 mm was observed after AM. One of the 22 Aa positive patients and 4 of 17 Pg positive patients became negative for these species after IT. The number of patients with detectable Pi decreased from 16 to 10 after IT. After AM, in comparison to baseline, suppression below detection level for Aa was achieved in 19 out of 22, for Pg in 9 out of 17, for Bf in 13 out of 14, and for Pi in 11 out of 16 patients. By contrast, higher frequencies of Peptostreptococcus micros and Fusobacterium nucleatum were found after AM. On the basis of the microbiological results the study group was separated into 2 subgroups: group A consisted of subjects who had no detectable levels of Aa, Pg, Bf and5% of Pi after AM. Group B consisted of those who still showed presence of one of these 3 species and/oror =5% levels of Pi. After AM, group B had significantly higher PI, BI, PPD and CAL scores then group A. It is concluded that group A showed low plaque scores and no detectable periodontal pathogens. This microbiological condition has been associated with a long-term stable periodontium.