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Notes: 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.

Notes: Background/aims: In an Indonesian population deprived of regular dental care, the experienced progression of disease between baseline (1987) and follow-up (1994) was investigated in relation to the composition of the subgingival microbiota at follow-up. At baseline the age ranged from 15 to 25 years. Clinical and microbiological evaluation was completed in 158 of the 167 subjects available at follow-up.Methods: Plaque index (PI), pocket depth (PD), bleeding on probing (BOP), and attachment loss (AL) were scored at the approximal surfaces of all teeth and subgingival calculus on the approximal surfaces of the Ramfjord teeth only (number of sites with subgingival calculus: NSC). A pooled sample of the deepest pocket in each quadrant was evaluated using microbiological culture techniques.Results: At baseline the mean values of the clinical parameters were AL=0.35 mm, PI=1.01, BOP=0.80 PD=3.25 mm and NSC=6.04 and at follow-up AL=0.75 mm, PI=1.16, BOP=1.19, PD=3.34 mm and NSC=5.85. All parameters except PD and NSC showed a statistically significant increase. At follow-up the prevalence of Actinobacillus actinomycetemcomitans was 40%, of Porphyromonas gingivalis 67%, of Prevotella intermedia 66%, of Fusobacterium nucleatum 79%, of Bacteroides forsythus 16%, of Campylobacter rectus 4%, and of P. micros 6%. No differences in clinical parameters were found between groups with or without these micro-organisms. In 129 subjects AL of 〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:03036979:JCPE280703:ges" location="ges.gif"/〉2 mm at 〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:03036979:JCPE280703:ges" location="ges.gif"/〉1 site was found. Logistic regression showed three significant odds-ratio’s for experienced progressive periodontitis: Plaque index (12.2), gender (3.4) and Actinobacillus actinomycetemcomitans (2.9).Conclusions: The results of this retrospective study suggest that plaque is the most important parameter related to experienced disease progression, and that the presence of A. actinomycetemcomitans may be associated with increased chance of disease progression.

Notes: Abstract Background, aims: The aim of this double-blind, parallel study was to evaluate the adjunctive effects of systemically administered amoxicillin and metronidazole in a group of adult periodontitis patients who also received supra- and subgingival debridement.Methods: 49 patients with a diagnosis of generalised severe periodontitis participated in the study. Random assignment resulted in 26 patients in the placebo (P) group with a mean age of 40 years and 23 patients in the test (T) group which had a mean age of 45 years. Clinical measurements and microbiological assessments were taken at baseline and 3 months after completion of initial periodontal therapy with additional placebo or antibiotic treatment. Patients received coded study medication of either 375 mg amoxicillin in combination with 250 mg metronidazole or identical placebo tablets, every 8 hours for the following 7 days.Results: At baseline, no statistically significant differences between groups were found for any of the clinical parameters. Except for the plaque, there was a significantly larger change in the bleeding, probing pocket depth (PPD) and clinical attachment level (CAL) in the T-group as compared to the P-group after therapy. The greatest reduction in PPD was found at sites with initial PPD of 〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:03036979:JCPE280402:ges" location="ges.gif"/〉7 mm, 2.5 mm in the P-group and 3.2 mm in the T-group. The improvement in CAL was most pronounced in the PPD category 〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:03036979:JCPE280402:ges" location="ges.gif"/〉7 mm and amounted to 1.5 mm and 2.0 mm in the P- and T-groups, respectively. No significant decrease was found in the number of patients positive for any of the test species in the P-group. The number of patients positive for Porphyromonas gingivalis, Bacteroides forsythus and Prevotella intermedia in the T-group showed a significant decrease. After therapy there was a significant difference between the P- and the T- group in the remaining number of patients positive for P. gingivalis, B. forsythus and Peptostreptococcus micros. 4 subgroups were created on the basis of the initial microbiological status for P. gingivalis positive (Pg-pos) and negative patients (Pg-neg) in the P- and the T-groups. The difference in reduction of PPD between Pg-pos and Pg-neg patients was particularly evident with respect to the changes in % of sites with a probing pocket depth 〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:03036979:JCPE280402:ges" location="ges.gif"/〉5 mm. This % decreased from 45% at baseline to 23% after treatment in the Pg-pos placebo subgroup and decreased from 46% to 11% in the Pg-pos test subgroup (p〈inlineGraphic alt="leqslant R: less-than-or-eq, slant" extraInfo="nonStandardEntity" href="urn:x-wiley:03036979:JCPE280402:les" location="les.gif"/〉0.005). In contrast, the changes in the proportions of sites with a probing pocket depth 〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:03036979:JCPE280402:ges" location="ges.gif"/〉5 mm in the Pg-neg placebo and Pg-neg test subgroup were similar, from 43% at baseline to 18% after treatment versus 40% to 12%, respectively.Conclusions: This study has shown that systemic usage of metronidazole and amoxicillin, when used in conjunction with initial periodontal treatment in adult periodontitis patients, achieves significantly better clinical and microbiological results than initial periodontal treatment alone. Moreover, this research suggests that especially patients diagnosed with P. gingivalis benefit from antibiotic treatment.

Notes: 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.

Notes: Background: The widespread use of antibiotics for prophylaxis and treatment of bacterial infections has lead to the emergence of resistant human pathogens. Great differences have been documented between European countries in the use of systemic antibiotics. In parallel, significant differences in levels of resistant pathogens have been documented.〈section xml:id="abs1-2"〉〈title type="main"〉Aim:To investigate whether differences in antibiotic use influence the level of antimicrobial resistance of the subgingival microflora of untreated patients with adult periodontitis in The Netherlands and Spain.Method: Blood agar plates containing breakpoint concentrations of penicillin, amoxicillin, amoxicillin and clavunalate, metronidazole, erythromycin, azithromycin, clindamycin and tetracycline were used to determine the proportion of bacteria from the subgingival plaque that was resistant to these antibiotics. In the Spanish patients, statistically significant higher mean levels of resistance were found for penicillin, amoxicillin, metronidazole, clindamycin and tetracycline. The mean number of different bacterial species growing on the selective plates was higher in the Spanish patients, as was the % of resistant strains of most periodontal pathogens. A striking difference was observed in the frequency of occurrence of tetracycline-resistant periodontal pathogens. In Spain, 5 patients had 〈inlineGraphic alt="geqslant R: gt-or-equal, slanted" extraInfo="nonStandardEntity" href="urn:x-wiley:03036979:JCPE270201:ges" location="ges.gif"/〉3 tetracycline resistant periodontal pathogens, whereas this was not observed in any of the Dutch patients.Conclusions: The widespread use of antibiotics in Spain is reflected in the level of resistance of the subgingival microflora of adult patients with periodontitis.

Notes: Aim: This study evaluated the microbial effects of a newly formulated mouthwash (Halita®) 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.

Notes: 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〈0.005). In the placebo group, no significant reduction in the mean organoleptic score occurred. Consequently, this resulted, after 2 weeks, in a greater change of the organoleptic scores in the test group in comparison to the placebo group (p〈0.005). The mean VSC scores were reduced from 292 to 172 ppb in the test group (p〈0.005), whereas no reduction was observed in the placebo group. At the 2-week examination, the mean change of the VSC scores in the test group was significantly greater than the mean change in the placebo group (p〈0.005). Neither in the test nor in the placebo group a significant reduction in tongue coating was observed.Conclusions:  In conclusion, the tested mouthrinse is effective in the treatment of oral halitosis.

Notes: 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.

Notes: 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.

Notes: Objectives: The CARD15 gene encodes the Nod2 protein, which is involved in intracellular recognition of bacterial products like peptidoglycan, activates inflammation and regulates apoptosis through nuclear factor-kappa B, a transcription factor that plays a central role in the innate immunity. Two functional mutations, an insertion mutation at nucleotide 3020 (3020insC) and a missense mutation C2104T in the CARD15 gene (originally NOD2 gene) have been reported to be associated with Crohn's disease. Our aim was to investigate the occurrence of CARD15 gene polymorphisms in adult patients with periodontitis taking into account smoking and presence of putative periodontal pathogens as additional variables.Material and methods: A case–control study was performed in 104 Dutch Caucasian patients with severe adult periodontitis (54 non-smokers and 50 smokers, mean age 46 years) and in 97 ethnically matched, periodontal healthy controls (73 non-smokers and 24 smokers, mean age 40 years). DNA isolated from a mouthwash was typed with PCR technology. Presence of putative periodontal pathogens was established by culture technique.Results: Frequencies of the CARD15 3020insC and 2104T mutations were similar in the periodontitis group and in the control group (5.1% and 13.3%; 5.2% and 10.3%, respectively). The highest carrier frequency of CARD15 mutations was found in non-smoking patients without Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans (29.4% versus 17.4% in controls); however it did not reach statistical significance.Conclusion: Our results suggest no role for CARD15 3020insC and C2104T mutations in adult periodontitis.