Susan Brailsford

The Predominant Microflora of Nursing Caries Lesions

The predominant microflora recovered from infected dentine of 52 carious teeth from 14 children with nursing caries was determined using both selective and non–selective media for the isolation of specific genera and acidified media (pH 5.2) to isolate the predominant aciduric microorganisms, and compared with the microflora of sound enamel surfaces in caries–free children. Streptococcus mutans formed a significantly greater proportion of the lesion flora while Streptococcus oralis, Streptococcus sanguis and Streptococcus gordonii formed a significantly greater proportion of the plaque flora from sound tooth surfaces. The proportions of Actinomyces naeslundii and Actinomyces odontolyticus were significantly greater in the plaque samples than in the lesion samples. Actinomyces israelii formed 18.2% of the flora from the lesions, but was not isolated from the plaque samples. The proportions of Candida albicans, Lactobacillus spp. and Veillonella spp. were also significantly greater in the carious dentine than in the plaque samples. The most frequently isolated lactobacilli were Lactobacillus casei, Lactobacillus fermentum and Lactobacillus rhamnosus. The predominant aciduric flora was S. oralis, S. mutans and A. israelii and these taxa were also isolated from a similar proportion of the lesions at pH 7.0. Strains of S. mutans, L. casei, L. fermentum and L. rhamnosus isolated from individual carious teeth were genotyped using PCR–based methods. Each species was genotypically heterogeneous and different genotypes were recovered from different carious teeth in the same child. These data indicate that the microflora of lesions in the same child is microbiologically diverse and support a non–specific aetiology for nursing caries in which the physiological characteristics of the infecting flora, not its composition, is the major determinant underlying the disease process.

Comparison between Visual Examination and a Laser Fluorescence System for in vivo Diagnosis of Occlusal Caries

This study compared a laser fluorescence (LF) system (DIAGNOdent) with a visual caries scoring system for in vivo detection and diagnosis of occlusal caries under the conditions of an epidemiological study, in 132 mandibular and 38 maxillary first permanent molars in 170 children (mean age: 6.85 ± 0.58 years). The teeth were cleaned and occlusal caries status in a selected investigation site recorded using both visual and LF systems. The LF readings were interpreted both according to the cut–off points recommended by the manufacturers and those based on laboratory research with histological validation. The percentage agreement of the LF and visual scoring system was better using the cut–off limits recommended by the manufacturer. Histological validation was not possible in this clinical study, but it appeared that either the LF method was overscoring some lesions or the visual method was underscoring them. Since the LF instrument cannot be expected to differentiate caries from hypomineralisation, it should probably be used as an adjunct to a clinical examination.

Phenotypic and Genotypic Selection of Microbiota Surviving under Dental Restorations

ABSTRACT The effects of sealing infected carious dentine below dental restorations on the phenotypic and genotypic diversity of the surviving microbiota was investigated. It was hypothesized that the microbiota would be subject to nutrient limitation or nutrient simplification, as it would no longer have access to dietary components or salivary secretion for growth. The available nutrients would be limited primarily to serum proteins passing from the pulp through the patent dentinal tubules to the infected dentine. Ten lesions were treated, and infected dentine was sealed below dental restorations for approximately 5 months. Duplicate standardized samples of infected dentine were taken at baseline and after the removal of the restorations. The baseline microbiota were composed primarily of Lactobacillus spp., Streptococcus mutans, Streptococcus parasanguinis, Actinomyces israelii, and Actinomyces gerencseriae. None of these taxa were isolated among the microbiota of the dentine samples taken after 5 months, which consisted of only Actinomyces naeslundii, Streptococcus oralis, Streptococcus intermedius, and Streptococcus mitis. The microbiota of the final sample exhibited a significantly (P < 0.001) increased ability to produce glycosidic enzymes (sialidase, β-N-acetylglucosaminidase, and β-galactosidase), which liberate sugars from glycoproteins. The genotypic diversity of S. oralis and A. naeslundii was significantly (P = 0.002 and P = 0.001, respectively) reduced in the final samples. There was significantly (P < 0.001) greater genotypic diversity within these taxa between the pairs of dentine samples taken at baseline than was found in the 5-month samples, indicating that the dentine was more homogenous than it was at baseline. We propose that during the interval between placement of the restorations and their removal, the available nutrient, primarily serum proteins, or the relative simplicity and homogeneity of the nutrient supply significantly affected the surviving microbiota. The surviving microbiota was less complex, based on compositional, phenotypic, and genotypic analyses, than that isolated from carious lesions which were also exposed to salivary secretions and pH perturbations.

Dental Caries, Oral Hygiene, and Oral Clearance in Children with Craniofacial Disorders

The reason that children with cleft palates tend to have a greater prevalence of tooth decay than normal children is unclear. We hypothesized that children with cleft palates would have increased oral clearance times for foods and, consequently, higher levels of caries and caries-associated micro-organisms than control children. Children aged 6–16 yrs, with (n = 81) or without (n = 61) cleft palates, were studied. Children with cleft palates had DMFT and dmft scores greater (p < 0.01) than those of the control group. The number of caries-associated organisms was greater in the saliva of the cleft palate children (all p < 0.001). The oral hygiene, plaque and gingival index scores were greater (p < 0.0001), oral clearance was longer (p < 0.01), and levels of sucrose and starch-derived saccharides higher (p < 0.01) in the cleft palate group. However, salivary concentrations of organic acids were lower in the children with craniofacial disorders, probably reflecting the altered physiology of the more mature dental biofilm. The longer oral clearance times of foods and the consequent generation of fermentable sugars from starches may contribute to the higher caries prevalence observed in children with cleft palates.

Genotypic Heterogeneity of Streptococcus oralis and Distinct Aciduric Subpopulations in Human Dental Plaque

ABSTRACT The genotypic heterogeneity of Streptococcus oralisisolated from the oral cavity was investigated using repetitive extragenic palindromic PCR. Unrelated subjects harbored unique genotypes, with numerous genotypes being isolated from an individual.S. oralis is the predominant aciduric bacterium isolated from noncarious tooth sites. Genotypic comparison of the aciduric populations isolated at pH 5.2 with those isolated from mitis-salivarius agar (MSA) (pH 7.0) indicated that the aciduric populations were genotypically distinct in the majority of subjects (χ2 = 13.09; P = 0.0031). Neither the aciduric nor the MSA-isolated strains were stable, with no strains isolated at baseline being isolated 4 or 12 weeks later in the majority of subjects. The basis of this instability is unknown but is similar to that reported for Streptococcus mitis. Examination ofS. oralis strains isolated from cohabiting couples demonstrated that in three of five couples, genotypically identical strains were isolated from both partners and this was confirmed by using Salmonella enteritidis repetitive element PCR and enterobacterial PCR typing. These data provide further evidence of the physiological and genotypic heterogeneity of non-mutans streptococci. The demonstration of distinct aciduric populations of S. oralis implies that the role of these and other non-mutans streptococci in the caries process requires reevaluation.

The effects of the combination of chlorhexidine/thymol- and fluoride-containing varnishes on the severity of root caries lesions in frail institutionalised elderly people

Abstract Objectives. To compare the clinical effects of a fluoride-containing varnish (Fluor-Protector) in combination with a chlorhexidine-containing varnish (Cervitec) on existing root caries lesions in a group of frail elderly subjects. Methods. A randomised double blind longitudinal study was utilised. Subjects (n=102) were randomly allocated to a Test or Placebo group. All leathery and soft root caries lesions in all subjects were coated with Fluor-Protector while the lesions in the Test group were also coated with Cervitec and the lesions in the Placebo group were coated with a Placebo varnish. Treatments were repeated five times in a 12-month period. Clinical parameters associated with root caries, measurements of individual lesions and salivary levels of caries associated bacteria were made at intervals. Results. The clinical severity of the lesions in the Test group did not change significantly during the 12-month study period. In the Placebo group the mean lesion width and lesion height and length of exposed root increased significantly and the lesions were significantly closer to the gingival margin. There were no significant changes in the salivary levels of caries-associated microorganisms after 12 months although, in both groups, there was initially a significant reduction in the salivary levels of mutans streptococci. Conclusions. The combination of Fluor-Protector and Cervitec is a useful, simple, quick and non-invasive method for the control and management of existing root caries lesions. The procedure could be performed by a dental hygienist and may be usefully applied in other high-risk groups including persons with Parkinsons disease, debilitating neuromuscular conditions and dry mouth from whatever cause.

Oral candidosis in community-based patients with advanced cancer

Oral candidosis is a significant cause of morbidity in patients with advanced cancer. The objectives of the study were to determine the epidemiology, etiology, clinical features, and microbiological aspects of oral candidosis among community-based patients. The subjects were recruited from hospice day centers in England. The clinical component involved completion of a study questionnaire, assessment of performance status, clinical examination of the oral cavity, measurement of unstimulated whole salivary flow rate, measurement of stimulated whole salivary flow rate, and performance of an oral rinse. Oral swabs were taken from subjects with appearances indicative of oral candidosis. The laboratory component involved standard procedures to isolate, enumerate, and identify yeasts from the clinical specimens. Three hundred ninety patients participated in the study. Two hundred seventy-two (70%) patients had oral yeast carriage, and 51 (13%) patients had combined clinical and microbiological evidence of oral candidosis. On univariate analysis, the presence of oral candidosis was associated with poor Eastern Cooperative Oncology Group performance status, presence of a denture, usage of a systemic corticosteroid, the severity of xerostomia, a low unstimulated whole salivary flow rate, and a low stimulated whole salivary flow rate. Multivariate analysis identified poor Eastern Cooperative Oncology Group performance status, presence of a denture, usage of a systemic corticosteroid, and the severity of xerostomia as being independently associated with the presence of oral candidosis. Oral candidosis is relatively common in community-based patients with advanced cancer. Hence, such patients should be screened for oral candidosis and should also be screened for reversible factors that predispose to oral candidosis, such as poor dental hygiene and salivary gland dysfunction.

The microflora of the erupting first permanent molar

The relationship between microflora, eruption status and caries status in the first permanent molar of young children was investigated in 177 children aged 6–7 years. A significantly greater proportion of fully erupted teeth were classified as sound and plaque-free compared to partially erupted teeth. Fully erupted teeth yielded greater numbers and proportions of mutans streptococci compared with significantly greater numbers and proportions of Actinomyces israelii in partially erupted teeth. Logistical regression analysis showed significant associations between white spot lesions in partially erupted teeth and increased numbers of Streptococcus oralis, mutans streptococci and Streptococcus salivarius whereas the presence of Actinomyces naeslundii was associated with health. Significantly greater numbers and proportions of S. oralis and S. salivarius were isolated from partially erupted teeth with white spot lesions whereas Streptococcus mutans was isolated in significantly greater numbers and proportions from fully erupted molars with white spots. This study suggests that organisms other than mutans streptococci are associated with caries development in erupting permanent molar teeth.

Effect of the environment on genotypic diversity of Actinomyces naeslundii and Streptococcus oralis in the oral biofilm.

ABSTRACT The genotypic diversity of Actinomyces naeslundii genospecies 2 (424 isolates) and Streptococcus oralis (446 isolates) strains isolated from two sound approximal sites in all subjects who were either caries active (seven subjects) or caries free (seven subjects) was investigated by using the repetitive extragenic palindromic PCR. The plaque from the caries-active subjects harbored significantly greater proportions of mutans streptococci and lactobacilli and a smaller proportion of A. naeslundii organisms than the plaque sampled from the caries-free subjects. These data confirmed that the sites of the two groups of subjects were subjected to different environmental stresses, probably determined by the prevailing or fluctuating acidic pH values. We tested the hypothesis that the microfloras of the sites subjected to greater stresses (the plaque samples from the caries-active subjects) would exhibit reduced genotypic diversity since the sites would be less favorable. We found that the diversity of A. naeslundii strains did not change (χ2 = 0.68; P = 0.41) although the proportional representation of A. naeslundii was significantly reduced (P < 0.05). Conversely, the diversity of the S. oralis strains increased (χ2 = 11.71; P = 0.0006) and the proportional representation of S. oralis did not change. We propose that under these environmental conditions the diversity and number of niches within the oral biofilm that could be exploited by S. oralis increased, resulting in the increased genotypic diversity of this species. Apparently, A. naeslundii was not able to exploit the new niches since the prevailing conditions within the niches may have been deleterious and not supportive of its proliferation. These results suggest that environmental stress may modify a biofilm such that the diversity of the niches is increased and that these niches may be successfully exploited by some, but not necessarily all, members of the microbial community.

Effect of biofilm growth on expression of surface proteins of Actinomyces naeslundii genospecies 2

ABSTRACT The predominant surface proteins of biofilm and planktonic Actinomyces naeslundii, a primary colonizer of the tooth surface, were examined. Seventy-nine proteins (the products of 52 genes) were identified in biofilm cells, and 30 of these, including adhesins, chaperones, and stress-response proteins, were significantly up-regulated relative to planktonic cells.