Sulfate-reducing bacteria in relation with other potential periodontal pathogens.

BACKGROUND, AIMS: Oral sulfate-reducing bacteria are involved in several clinical categories of periodontitis. The aim of this cross-sectional study was to compare the presence of sulfate-reducing bacteria (SRB) with other putative pathogens including spirochetes, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis, and Treponema denticola in periodontal lesions. METHOD: Periodontal SRB were detected by enrichment culture and compared with a microscopic spirochete count (n=168). Species-specific oligonucleotide probes directed against the 16S rRNA were employed to determine the presence of A. actinomycetemcomitans, P. gingivalis, B. forsythus, and T. denticola (n=55). RESULTS: A significant positive correlation was observed between the presence of SRB and the proportions of spirochetes in subgingival plaque, although the 2 bacterial groups also occurred separately. SRB tended to be negatively correlated with the presence of A. actinomycetemcomitans. In contrast, all pockets with SRB harbored either T. denticola, or both T. denticola and B. forsythus (12/14) before therapy. Interestingly, the combination of SRB with P. gingivalis occurred in 32% of the periodontal pockets before treatment. After initial periodontal therapy, the prevalence of this combination was reduced to 2% of the sites, and to 25% of the sites in recall patients. CONCLUSION: The presence of SRB was positively correlated with T. denticola, B. forsythus, and P. gingivalis in periodontal lesions. These suspected pathogens form a complex strongly associated with destructive periodontitis.

Decrease of sulfate-reducing bacteria after initial periodontal treatment.

Sulfate-reducing bacteria (SRB) are associated with periodontitis, but it is unknown if elimination of these potential pathogens accompanies clinical improvement. This longitudinal study examined the occurrence of SRB and clinical effects following scaling and root planing. In this study, the presence of periodontal SRB was determined in 38 selected patients before and six months after mechanical therapy. SRB were detected by the enrichment culture technique. Mechanical periodontal treatment resulted in elimination of SRB in 89% of the patients, and 95% of the sites (n = 76). SRB were significantly reduced in patients with progressive, adult, and refractory periodontitis. The elimination of SRB was accompanied by clinical improvement. The mean gain of attachment of these pockets was 3 mm (p < 0.001). The reductions in pocket depth (p < 0.001) and bleeding were significant (p < 0.001). Persistence of SRB correlated with the initial pocket depth (p < 0.02) and attachment level (p < 0.02), and with bleeding of the site after treatment (p < 0.05). In conclusion, mechanical debridement is generally effective for the elimination of SRB.

Isolation of Desulfomicrobium orale sp. nov. and Desulfovibrio strain NY682, oral sulfate-reducing bacteria involved in human periodontal disease.

The species of sulfate-reducing bacteria that prevail in sites affected by periodontal disease may be different from those commonly occurring in the digestive tracts of healthy individuals. Ten strains of mesophilic sulfate-reducing bacteria (SRB) were isolated from subgingival plaque in periodontal lesions of ten patients with periodontitis. Characterization on the basis of morphological, physiological and phylogenetic properties demonstrated two distinct types of oral SRB. One strain was a curved rod with high motility. For dissimilatory sulfate reduction, lactate or pyruvate was oxidized incompletely to equimolar amounts of acetate. Desulfoviridin and cytochrome c3 were present in this mesophilic vibrio and the cellular lipid profile was similar to that from members of the genus Desulfovibrio. The 16S rDNA sequence was similar to that of the proposed 'Desulfovibrio fairfieldensis'. Cells of the nine other strains were straight, rod-shaped, exhibited a low growth rate and oxidized substrates incompletely to acetate. These SRB, like members of the genus Desulfomicrobium, lacked desulfoviridin. Analysis of the 16S rDNA sequences of seven of the nine isolates showed a high degree of similarity among these oral strains, forming a distinct lineage within the genus Desulfomicrobium. The cellular lipid profile of a representative oral strain, NY678T, was in accordance with that of other Desulfomicrobium species, but also showed dissimilar features. The phenotypic and phylogenetic analyses indicate that these rod-shaped SRB from the oral cavity could be regarded as a new species, for which the designation Desulfomicrobium orale sp. nov. is proposed.

Sulfate-reducing bacteria in association with human periodontitis.

BACKGROUND, AIMS: Sulfate-reducing bacteria (SRB) may be etiologically involved in destructive periodontal diseases. These strictly anaerobic bacteria utilize fermentation products for energy conservation by reduction of sulfate to sulfide. This toxic product can accumulate in periodontal pockets in concentrations causing cellular destruction. SRB depend on an actively degrading microbiota to produce a reduced environment, fermentation products and sulfate. The detection frequency of these bacteria is strongly increased in periodontitis compared with healthy sites in the oral cavity. METHOD: In this study, the presence of SRB was determined in relation to clinical features of the patients and to site-specific clinical parameters of periodontitis, such as pocket depth, bleeding and attachment level. Patients with clinical characteristics of severe periodontitis (n=87) were included in the study, 78 were untreated patients and 9 patients were in maintenance care after treatment. Samples were taken (n=261) from the deepest periodontal pockets, and presence of SRB was determined by enrichment culture in an anoxic chamber. RESULTS: In 64% of the patients, SRB were present in at least 1 pocket. They occurred among patients from 23 to 57 years old, and tended to prevail among patients older than 30 years. There was a tendency to increased SRB occurrence among patients with more than 50% of bleeding sites, or with several angular bony defects or furcations. In 44% of the periodontal pockets SRB were present. They tended to prevail in pockets showing bleeding on probing, furcations, angular bony defects, or an endodontal complication. Presence of SRB was positively correlated with increased pocket depth (p<0.05). SRB were found to be associated with various clinical categories of periodontitis, including early onset periodontitis, rapidly progressive periodontitis, adult periodontitis, and refractory periodontitis. Although SRB predominated among patients with an adult form of periodontitis, i.e., with an occurrence of 72%, there was no significant correlation with age of the patient. Among treated patients under maintenance care, SRB prevalence was significantly reduced in comparison with untreated patients (p<0.02). Occurrence of SRB in periodontal pockets showed an odds ratio of 11.2 in comparison with healthy oral sites. CONCLUSION: Periodontal sulfate-reducing bacteria are associated with several clinical categories of periodontitis and with periodontal sites of increased pocket depth.

Sulfate-reducing bacteria in periodontal pockets and in healthy oral sites.

The aim of this study was to monitor the presence of sulfate-reducing bacteria (SRB) at different sites in the mouths of both healthy individuals and periodontitis patients. In 20 healthy subjects and 21 periodontitis patients, samples were taken from the palate, vestibulum, dorsum of the tongue, supragingival plaque, and periodontal pockets. In order to demonstrate growth of SRB, samples were incubated in an anoxic chamber in a reduced growth-medium for SRB, with an iron-indicator for sulfide production. The SRB were detected throughout the oral cavity. They were found on the mucosa in 10% of both healthy subjects and periodontitis patients. On the tongue and in supragingival plaque, the frequency of detection was slightly higher (22% of the subjects). In contrast, 86% of the periodontitis patients harbored SRB in one or more pockets. In 1/3 of the patients, SRB were present in all 3 pockets that were sampled. The data indicated that SRB belong to the normal oral microbiota, and have a preference for periodontal pockets.

Proteolysis and utilization of albumin by enrichment cultures of subgingival microbiota.

Subgingival dental plaque consists mainly of microorganisms that derive their energy from amino acid fermentation. Their nutrient requirements are met by the subgingival proteolytic system, which includes proteases from microorganism and inflammatory cells, and substrate proteins from sulcus exudate, including albumin. To determine the selective effect of individual proteins on microbiota, we used albumin as the main substrate for growth. Eight subgingval plaque samples from untreated periodontal pockets of patients with adult periodontitis were inoculated in peptone yeast medium with bovine albumin (9 g/l). After three subculture steps, cell yields of the enrichment cultures at the medium with 0, 1.25, 2.5, 5, 10, and 20 g/l albumin were determined. Proteolytic activity (U/absorbance at 550 nm) of the enrichment cultures and different isolates derived from the cultures was estimated by the degradation of resorufin-labeled casein. It was observed that the yield of the mixed culture was albumin limited, and the proteolytic activities of the cultures in albumin broth were higher than in control (peptone broth). Among the isolates from the enrichment cultures, Peptostreptococcus micros, Prevotella melaninogenica, Prevotella buccae and Prevotella bivia demonstrated proteolysis. The frequent occurrence of Streptococcus gordonii and Streptococcus anginosus in the albumin cultures is explained by their ability to utilize arginine as an energy source for growth. Albumin in the medium was partly degraded by pure cultures but completely consumed in enrichment cultures, indicating synergy of bacterial proteinases. It is concluded that the subgingival microbiota possesses proteolytic activity and may use albumin as a substrate for their growth. Enrichment cultures on albumin may serve as a relatively simple in vitro model to evaluate the effects of proteinase inhibitors.

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.

The importance of immunoglobulin-breakdown supporting the growth of bacteria in oral abscesses.

Oral bacteria play an important rôle in the causation of oro-facial abscesses. However, they can also be involved in brain, liver and lung abscesses. To persist, it is essential that these bacteria can grow on those sites. The main source of nutrients for growth in abscesses is likely to be tissue exudate, which is rich in serum-derived proteins, and relatively poor in free amino acids and carbohydrates. Degradation of intact proteins seems a crucial step in providing the peptides necessary for energy generation. The aim of this study was to investigate the capacity of microorganisms from asscesses to degrade serum proteins, in particular immunoglobulins. To this end, samples were taken by aspiration from 16 odontogenic abscesses. It was found that pus from abscesses differed strongly in the concentration of viable bacterial cells. The ability of the abscess microflora to degrade serum proteins was investigated after growth of the sample in heat-inactivated human serum. The microflora from abscesses with a high concentration (n = 10) of bacteria strongly degraded immunoglobulins, whereas breakdown of immunoglobulins was virtually absent after growth of the microflora from low-bacterial concentration (n = 6) abscesses. Bacteriological analyses revealed the presence of at least one proteinase-producing species, like Porphyromonas, black-pigmented Prevotella species, or Actinomyces meyeri, in abscesses with a high density of bacteria, but not in those with low bacterial density. The results indicate that the capacity to degrade intact proteins, in particular immunoglobulins, is a major determinant of bacterial growth in abscesses.

The effect of mouthrinses containing zinc and triclosan on plaque accumulation, development of gingivitis and formation of calculus in a 28-week clinical test.

Experimental mouthrinses containing 0.4% zinc sulphate and 0.15% triclosan, which differed in base formulations were compared to a commercially available non-active control mouthrinse. Following baseline clinical examinations for plaque, gingival bleeding and calculus, the volunteers were provided with a dental prophylaxis and given oral hygiene instruction, stratified into 3 groups and given 1 of 3 mouthrinses. Further clinical assessments were performed after 4, 16 and 28 weeks. Salivary mutans streptococci were also monitored during the study. At 4 weeks, plaque and calculus scores in all groups were low compared to baseline. During the remainder of the study, these improvements were not maintained and both plaque and calculus levels increased in all groups. Plaque was significantly lower (P < 0.05) than in the control at all time points. Calculus was significantly lower (P < 0.05) than in the control at all time points. Calculus was significantly lower at week 28 for experimental mouthrinse group 2. Gingival bleeding also decreased in the initial 4 weeks but increased thereafter in the control group. In contrast, gingival bleeding was significantly (P < 0.05) lower in the two experimental groups than in the control group. No significant changes in mutans streptococci were observed.

Effect of chlorhexidine varnish on Actinomyces naeslundii genospecies in plaque from dental fissures.

This study describes the effects of varnish containing 40% chlorhexidine diacetate on Actinomyces naeslundii populations in plaque from human molar fissures. In each of 15 subjects two dental fissures with high levels of mutants streptococci were selected. The experimental treatment consisted of the single application of a small amount of chlorhexidine varnish onto the selected fissures. The varnish was removed 15 min after application. One month after varnish application a significant increase was observed in A. naeslundii counts while the number of mutans streptococci had decreased significantly compared with preexperimental levels. From 85 randomly selected Actinomyces isolates taken from blood agar plates before varnish application, 44% belonged to A. naeslundii genospecies 1 and 56% to A. naeslundii genospecies 2. From 106 isolates taken 1 month after chlorhexidine varnish application, 42% belonged to A. naeslundii genospecies 1 and 58% to A. naeslundii genospecies 2. At baseline 28% of A. naeslundii genospecies 1 strains were catalase-positive, but 1 month after varnish application 4% of the strains were catalase-positive (p < 0.05). It is concluded that chlorhexidine varnish application caused an increase of A. naeslundii in dental plaque, but induced no significant changes in the distribution of the two A. naeslundii genospecies.

Cariogenicity depends more on diet than the prevailing mutans streptococcal species.

This review aims to compare the occurrence and distribution of mutans streptococci in Africa, Europe, and North America and in addition will try to offer explanations for existing relationships among salivary mutans streptococci counts, dietary patterns, and dental caries. The literature reveals that salivary mutans streptococci counts in child populations of the three continents are comparable. The distribution of mutans streptococci species, with a predominance of S. mutans followed by S. sobrinus, and the virtual absence of other mutans streptococci species are also comparable. Although it is widely believed that diet has an important effect on mutans streptococci counts, this review provides evidence that this does not hold true when variations in dietary patterns are moderate, as they normally are in real-life situations. Since the diets of the child populations in the three continents vary moderately, a strong dietary-induced effect on salivary mutans streptococci counts cannot be expected. The observed analogous salivary mutans streptococci counts in these child populations are thus 'not surprising' but are in accordance with the conceptual expectation. The differences in caries experience in children of the three continents cannot be explained by the prevailing mutans streptococci species but instead should be attributed to differences in the cariogenicity of the various diets. The fact that the cariogenicity of the diet determines the development of dental caries while hardly affecting the mutans streptococci counts explains the limited value of the latter as an indicator of dental caries. The reviewed literature shows that mutans streptococci are ubiquitous in children aged 7 years and older in Africa, Europe, and North America. Mutans streptococci should therefore be considered as belonging to the indigenous microflora of the human mouth.

Sulfate-reducing bacteria in the periodontal pocket.

This report is the first to describe the occurrence of sulfate-reducing bacteria in the human mouth. Samples of subgingival dental plaque were examined for the presence of sulfate-reducing bacteria. Using enrichment cultures, sulfate-reducing bacteria were detected in 25 (58%) of 43 individuals, and in 39 (48%) of the 82 samples. Pure isolates of sulfate-reducing bacteria, obtained from a limited number of enrichment cultures, belonged to the genera Desulfobacter and Desulfovibrio. These genera are also the predominant sulfate-reducing bacteria in the human large intestine. The sulfate-reducing bacteria use sulfate as terminal electron acceptor to oxidize low-molecular-weight organic compounds, mainly products of microbial fermentation such as acetate, lactate etc. The numbers of sulfate-reducing bacteria in the mouth are assumed to be limited by sulfate. Potential sources of sulfate in the subgingival area include free sulfate in pocket fluid and glycosaminoglycans from periodontal tissues.

Characterization of immunoglobulin G-degrading proteases of Prevotella intermedia and Prevotella nigrescens.

Degradation of immunoglobulins is thought to be an important factor in the causation of periodontal diseases by hindering local host defenses and by providing nutrients to the periodontal microflora. In this study, we characterized the proteolytic activity against human immunoglobulin G (IgG) of 20 strains of Prevotella intermedia and Prevotella nigrescens isolated from periodontal pockets and oral abscesses. IgG degradation was studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. All strains degraded IgG within 48 h after growth in trypticase-yeast extract medium (TY) supplemented with 0.3% IgG. Incorporating IgG in TY broth enhanced bacterial growth. Protease profiles (zymography), which revealed the presence of 1-4 IgG-degrading proteolytic bands in bacterial cell extracts, became more complex after growth in the presence of IgG. A 38-kDa protease capable of degrading IgG nonspecifically was present in almost all strains. The proteolytic activity was mainly located on the surface of the cell envelope. Two strains of P. intermedia and P. nigrescens ATCC 33563 were selected for further studies. Bacterial cell suspensions in phosphate-buffered saline completely degraded human IgG, IgA and IgM within 24 h. This activity depended on reducing conditions and was inhibited at temperatures above 50 degrees C. The pH optimum of immunoglobulin degradation was at pH 7. Strains cultured at 42 degrees C showed a markedly reduced capacity to degrade IgG. Inhibition studies revealed that breakdown of IgG was caused by a cysteine protease(s). The capacity of P. intermedia and P. nigrescens to degrade immunoglobulins may explain their association with polymicrobial oral diseases.

Streptococci and actinomyces inhibit regrowth of Streptococcus mutans on gnotobiotic rat molar teeth after chlorhexidine varnish treatment.

Clinical studies suggest that the long-term suppression of mutans streptococci on tooth surfaces after intensive chlorhexidine therapy is mainly due to bacterial interference. Other streptococci and also Actinomyces naeslundii are proposed to inhibit regrowth of mutans streptococci after suppression by the agent. We have tested this hypothesis in gnotobiotic rats associated with Streptococcus mutans alone, or associated with S. mutans and strains of Streptococcus oralis, Streptococcus sanguis, Streptococcus gordonii, Streptococcus mitis biovar I, and A. naeslundii. Left lower jaws in these rats were treated with concentrated chlorhexidine varnish, and the return of S. mutans on the treated jaws monitored. In mono-associated rats, S. mutans regained the level of the untreated right lower jaw in approximately 1 week. In contrast, S. mutans remained suppressed for several weeks in rats multi-associated with other streptococci and actinomyces strains. The suppression was more pronounced in the rats fed on basal diet with little free sugars than in rats fed on a sucrose-containing diet. Counts of other streptococci recovered quickly from the intensive chlorhexidine treatment, but A. naeslundii remained suppressed for at least 1 week. The findings demonstrate the crucial importance of the oral microflora in controlling regrowth of mutans streptococci after chemotherapy.

Lactobacilli, mutants streptococci and dental caries: a longitudinal study in 2-year-old children up to the age of 5 years.

A 3-year cohort study was carried out in 252 pre-school children for early identification of caries-active individuals. During this period information was collected about the acquisition of mutants streptococci and lactobacilli from the age of 2 till 5 years old. At baseline mutants streptococci were detected in 43% of the children while the detection frequency of lactobacilli was low (11.5%). On an individual level, numbers of colony-forming units of mutans streptococci and lactobacilli in plaque and saliva varied largely during the study period. The correlations between the numbers of lactobacilli and mutants streptococci in the saliva of the mother and the saliva and plaque of the child were low and never exceeded r = 0.22. Very low correlations (< r = 0.22) were also found between the numbers of mutans streptococci or lactobacilli and the diet in terms of the number of sugar intakes. Nevertheless, in children older than 2.5 years correlations between the clinical caries score and lactobacilli in saliva (range 0.31-0.62) and mutans streptococci in plaque or saliva (range 0.24-0.46) were highly significant (p < 0.01).

Degradation of immunoglobulin G by periodontal bacteria.

Several subgingival microorganisms were tested for their ability to utilize human immunoglobulin G (IgG) as a substrate for growth. This was done using a protein-free chemically defined medium, supplemented with IgG. Stimulation of growth was observed for Capnocytophaga ochracea, Porphyromonas asaccharolytica, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella oralis, Lactobacillus catenaforme and Streptococcus intermedius. Immunoelectrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a protein assay demonstrated that P. intermedia and P. endodontalis completely degraded the protein chains of IgG. Partial breakdown of IgG was observed for P. asaccharolytica and C. ochracea, whereas P. oralis cleaved the IgG heavy chain, yielding Fc and Fab fragments. All these bacteria utilized IgG as a substrate for growth. Binding studies using an enzyme-linked immunosorbent assay, revealed complete loss of in vitro antigen-antibody binding capacity after incubation of specific IgG with P. endodontalis and partial loss of binding with P. intermedia, P. gingivalis, C. ochracea or Fusobacterium nucleatum. Degradation or inactivation of IgG by oral bacteria is thought to be important in the causation of polymicrobial infections.

The effect of mouthrinses containing zinc and triclosan on plaque accumulation and development of gingivitis in a 3-week clinical test.

Experimental mouthrinses containing 0.4% zinc sulphate and 0.15% triclosan were compared with a chlorhexidine and a negative control mouthrinse in a 3-week clinical trial. The zinc/triclosan mouthrinses 1 and 2 differed in their ethanol and humectant contents used to deliver the triclosan. The experimental protocol employed the partial mouth gingivitis design, whereby participants wear a toothshield during toothbrushing. Gingival health at baseline was established by professional cleaning, oral hygiene instruction and effective toothbrushing 3 x per day during a pre-experimental period of 2 weeks. The mouthrinses were subsequently used 2x daily following normal toothbrushing during 3 weeks. The pre-experimental oral hygiene phase very effectively reduced plaque levels and gingival bleeding. During the rinsing period, in the absence of mechanical removal of plaque from the protected teeth, gingival bleeding rose to above the prestudy level in the negative control group. The increments (change from baseline to 21 days) of plaque and bleeding scores for the zinc/triclosan mouthrinse 1 were significantly lower than those in the negative control group. As expected, plaque and gingivitis scores were lowest in the group that rinsed with chlorhexidine. The results extend previous observations on the efficacy of the zinc/triclosan system to maintain gingival health.

[The use of chlorhexidine varnish in the prevention of dental caries]. / Utilisation du vernis à la chlorhexidine dans la prévention des caries.

Dentistery recently has developed a new preventive treatment, that is the suppression of the acid-producing bacteria in dental plaque. The background and application of two chlorine hexidine lacquers, EC40 and Clorzoin are discussed.

Effect of chlorhexidine varnish on streptococci in dental plaque from occlusal fissures.

The treatment of tooth surfaces with chlorhexidine varnish may lead to long-lasting suppression of mutants streptococci in dental plaque. Microbiological observations following varnish treatment suggest that this prolonged suppression might be caused by bacterial interference. To investigate whether physiologically related organisms, such as other Streptococcus species, compete with mutans streptococci in the ecosystem, we have analyzed streptococcal populations on the tooth surface before and after chlorhexidine varnish treatment. Occlusal surfaces with high numbers of mutans streptococci were selected in human volunteers and treated with chlorhexidine varnish. Analyses of sequentially collected plaque samples confirmed that S. oralis-group streptococci returned to baseline levels shortly after the chlorhexidine application, while Actinomyces naeslundii populations reached prestudy or even higher levels only several days after treatment. Mutans streptococci, however, were below the detection level in the 14-day samples, except in 1 individual. The pattern of recolonization by individual Streptococcus species after chlorhexidine application closely resembled that of cleaned enamel surfaces: S. oralis and S. sanguis were primary colonizers while S. gordonii became dominant at a later stage. It is concluded that after intensive chlorhexidine treatment, a normal oral microflora reestablished, characterized by low proportions of mutans streptococci.

The use of lectins in monitoring degradation of oligosaccharide chains in mucin by oral streptococci.

The ability of utilize mucin oligosaccharides as sources of carbohydrate and energy is believed to be an important mechanism in the ecology of oral streptococci. In this study we have used digoxigenin-labelled lectins of various specificities to monitor changes in the nonreducing end groups of oligosaccharide chains following their degradation by Streptococcus oralis Ny 586 and Streptococcus sanguis Ny 584. The reaction of degraded mucin with peanut lectin, that recognizes the core disaccharide Gal (1,3)GalNAc in O-glycans, revealed a more extensive degradation of oligosaccharide by S. oralis than by S. sanguis. This corresponds to better growth of S. oralis on the mucin. Analyses with Datura stramonium lectin showed that terminal Gal (1,4)GlcNAc, or GlcNAc (1,4)GlcNAc moieties, in the oligosaccharides are attacked by both strains. Reaction patterns with alpha-L-fucose-specific lectins indicated that terminal fucose was released by S. oralis but not by S. sanguis. This was in accordance with sugar analyses which showed that approximately 40% of the fucose units were released. The results extend previously observed losses of sugars from oligosaccharide chains during growth of these organisms on mucin.