P. S. Bird

Oral Colonization of Streptococcus mutans in Six-month-old Predentate Infants

We hypothesize that S. mutans colonization occurs more frequently in pre-term children due to their relative immaturity. In this study of 172 predentate, six-month-old infants, we found that 50% of pre-term and 60% of full-term children harbored S. mutans. The colonization was confirmed by repeat sampling. Although there were minor differences, factors associated with S. mutans infection in pre-term and full-term infants were generally similar. In both groups, increased frequency of sugar was ranked the most important factor (p < 0.001), followed by breast-feeding (p < 0.001), and habits which allowed saliva transfer from mother to infant (p < 0.01). By contrast, non-colonization of S. mutans was associated with multiple courses of antibiotics (p < 0.001). Compared with pre-term children, there were higher percentages of full-term who had night feedings and consumed sugar during sleep times. Mothers with infected infants had S. mutans levels > 5 x 105 CFU/mL saliva (p < 0.001), poorer oral hygiene, more periodontal disease, and lower socio-economic status (p < 0.02) and snacked frequently (p < 0.001), compared with mothers with non-infected infants.

A Longitudinal Study of Streptococcus mutans Colonization in Infants after Tooth Eruption

We previously reported that, before tooth eruption, over one-half of infants aged 6 mos were already infected with Streptococcus mutans. The aim of this investigation was to determine the colonization of S. mutans after tooth eruption in the same cohort of 111 infants (35 pre-term, 76 full-term). Our results showed that S. mutans colonization increased with increasing age, so that by 24 mos of age, 84% harbored the bacteria (p < 0.01). The mean and median ages of S. mutans colonization in dentate infants were 15.7 mos and 16.0 mos, respectively. Factors associated with S. mutans colonization were sweetened fluids taken to bed (p < 0.01), frequent sugar exposure (p < 0.03) and snacking (p < 0.03), sharing of foods with adults (p < 0.03), and maternal S. mutans levels of > 105 CFU/mL (p < 0.02). In contrast, non-colonization of S. mutans was associated with toothbrushing (p < 0.03) and multiple courses of antibiotics (p < 0.001). Analysis of our data establishes the timing of S. mutans colonization in children from birth to 24 mos of age.

Association of Streptococcus mutans Infection and Oral Developmental Nodules in Pre-dentate Infants

Since dental caries may present soon after tooth eruption, we hypothesized that colonization of Streptococcus mutans can occur in the predentate stages. In this study, we examined S. mutans colonization and its association with oral developmental nodules (Bohns nodules) in 60 pre-term and 128 full-term, three-month-old infants. Overall, S. mutans was cultured from 30% (56/188) of the infants, and oral developmental nodules were noted in 55% (103/188). Compared with the pre-term, full-term infants showed a higher prevalence of S. mutans (34% vs. 20%, p < 0.02) as well as developmental nodules (61% vs. 42%, p < 0.05). In both groups, S. mutans was positively associated with numbers of developmental nodules in a dose-response relationship (p < 0.001), and with maternal salivary levels of the bacteria (p = 0.03). The permanence of S. mutans infection was confirmed by repeat saliva sampling at 6 months of age. Our results thus showed that many infants have already acquired S. mutans at 3 months of age, prior to tooth eruption.

Disinfection of dental stone casts: Antimicrobial effects and physical property alterations

OBJECTIVES The purpose of this study was to evaluate the effectiveness of disinfecting solutions incorporated into dental stone casts against a standard and representative group of microorganisms and to note changes in the physical properties of the casts. METHODS Irreversible hydrocolloid impressions were contaminated individually with Escherichia coli, Staphylococcus aureus, Enterobacter cloacae, Pseudomonas aeruginosa, Klebsiella pneumoniae, Actinobacter calcoaceticus, Bacillus subtilis, Mycobacterium phlei and Candida albicans. Four readily available disinfecting solutions (glutaraldehyde, povidone-iodine, chlorhexidine and sodium hypochlorite) were added to the die stone mix used to pour up the impressions. The set cast surfaces were swabbed at 1 h and 24 h, the samples plated on agar and incubated at 37 degrees C for 24 h and 3 d for M. phlei. Subsequently, colony forming units were counted. The physical properties assessed were setting time, setting expansion, compressive strength, detail reproduction and delayed expansion of the stone. RESULTS Only glutaraldehyde and povidone-iodine killed all contaminating microorganisms within 1 h, while the 1:5 dilution of sodium hypochlorite solution was equally effective after 24 h. Two percent glutaraldehyde was the most effective disinfectant with the least adverse effects on the physical properties of the set cast. Although povidone-iodine caused a decrease in the compressive strength of the set cast, it can be considered to be a sound alternative. SIGNIFICANCE This study supports the concept of incorporating disinfectants into model stone as a standard operating procedure for impressions of unknown history and, most sensibly, all dental impressions.

Effect of Fusobacterium nucleatum on the T and B cell responses to Porphyromonas gingivalis in a mouse model

T cell cytokine profiles and specific serum antibody levels in five groups of BALB/c mice immunized with saline alone, viable Fusobacterium nucleatum ATCC 25586, viable Porphyromonas gingivalis ATCC 33277, F. nucleatum followed by P. gingivalis and P. gingivalis followed by F. nucleatum were determined. Splenic CD4 and CD8 cells were examined for intracytoplasmic interleukin (IL)‐4, interferon (IFN)‐gamma and IL‐10 by dual colour flow cytometry and the levels of serum anti‐F. nucleatum and anti‐P. gingivalis antibodies determined by an ELISA. Both Th1 and Th2 responses were demonstrated by all groups, and while there were slightly lower percentages of cytokine positive T cells in mice injected with F. nucleatum alone compared with the other groups immunized with bacteria, F. nucleatum had no effect on the T cell production of cytokines induced by P. gingivalis in the two groups immunized with both organisms. However, the percentages of cytokine positive CD8 cells were generally significantly higher than those of the CD4 cells. Mice immunized with F. nucleatum alone had high levels of serum anti‐F. nucleatum antibodies with very low levels of P. gingivalis antibodies, whereas mice injected with P. gingivalis alone produced anti‐P. gingivalis antibodies predominantly. Although the levels of anti‐F. nucleatum antibodies in mice injected with F. nucleatum followed by P. gingivalis were the same as in mice immunized with F. nucleatum alone, antibody levels to P. gingivalis were very low. In contrast, mice injected with P. gingivalis followed by F. nucleatum produced equal levels of both anti‐P. gingivalis and anti‐F. nucleatum antibodies, although at lower levels than the other three groups immunized with bacteria, respectively. Anti‐Actinobacillus actinomycetemcomitans, Bacteroides forsythus and Prevotella intermedia serum antibody levels were also determined and found to be negligible. In conclusion, F. nucleatum immunization does not affect the splenic T cell cytokine response to P. gingivalis. However, F. nucleatum immunization prior to that of P. gingivalis almost completely inhibited the production of anti‐P. gingivalis antibodies while P. gingivalis injection before F. nucleatum demonstrated a partial inhibitory effect by P. gingivalis on antibody production to F. nucleatum. The significance of these results with respect to human periodontal disease is difficult to determine. However, they may explain in part differing responses to P. gingivalis in different individuals who may or may not have had prior exposure to F. nucleatum. Finally, the results suggested that P. gingivalis and F. nucleatum do not induce the production of cross‐reactive antibodies to other oral microorganisms.

Treatment of root canal biofilms of Enterococcus faecalis with ozone gas and passive ultrasound activation.

INTRODUCTION Biofilms of resistant species such as Enterococcus faecalis pose a major challenge in the treatment of root canals with established periapical disease. This study examined the effects of gaseous ozone delivered into saline on biofilms of E. faecalis in root canals of extracted teeth with and without the use of passive ultrasonic agitation. METHODS Biofilms of E. faecalis were established over 14 days in 70 single roots that had undergone biomechanical preparation followed by gamma irradiation. The presence and purity of biofilms were confirmed using scanning electron microscopy and culture. Biofilms were treated with saline (negative control), 1% sodium hypochlorite for 120 seconds (positive control), ozone (140 ppm ozone in air at 2 L/min delivered into saline using a cannula for 120 seconds), saline with passive ultrasonic activation (70 kHz and 200 mW/cm(2) applied to an ISO 15 file held passively within the canal, for 120 seconds), and ozone followed immediately by ultrasonic agitation. After treatment, samples were taken from the biofilm and serially diluted for plate counting. RESULTS Analysis revealed that 1% sodium hypochlorite was the most effective disinfecting agent followed by ozone combined with ultrasonic agitation, ozone alone, and finally ultrasonic alone. CONCLUSIONS Although none of the treatment regimes were able to reliably render canals sterile under the conditions used, ozone gas delivered into irrigating fluids in the root canal may be useful as an adjunct for endodontic disinfection.

Persistent Colonization with Tannerella forsythensis and Loss of Attachment in Adolescents

Colonization with Tannerella forsythensis may characterize the conversion of periodontally healthy sites into diseased sites. This three-year study describes the prevalence of T. forsythensis and its relationship to clinical loss of attachment (LOA) in a group of adolescents considered at risk of developing early chronic periodontitis. Adolescents with (LOA+) and without (LOA-) loss of attachment were examined at baseline and 1.5 and 3 yrs subsequently. On each occasion, attachment loss was measured on selected teeth, and the presence of T. forsythensis in their subgingival plaque samples was determined by PCR. T. forsythensis prevalence in LOA+ subjects at baseline (64%) increased to 82% and 86% on subsequent examinations. In contrast, prevalence of T. forsythensis in LOA- subjects was always significantly lower (25%, 36%, and 32%, respectively). The odds of loss of attachment were 8.16 times greater in subjects infected with T. forsythensis at each examination. These results suggest that T. forsythensis is strongly associated with loss of attachment in this adolescent population.

Oral Streptococcus species in pre‐term and full‐term children – a longitudinal study

BACKGROUND Despite high clinical significance, the microbiology of the dental biofilm in young children remains poorly understood. AIM The aim of this longitudinal study was to investigate five Streptoccocus species commonly found in the oral biofilm of children, namely Streptococcus mutans, Streptococcus sobrinus, Streptococcus mitis, Streptococcus sanguinis, and Streptococcus salivarius to determine their relative numbers in caries-free pre-term children, and age-matched full-term controls. DESIGN Plaque and saliva samples were obtained from 15 pre-term children and 15 age-matched controls at ages 3, 6, 12, 18, and 24 months. A quantitative real-time PCR technique was used to determine the numbers of five species of Streptococcus using probes and primers specific for each bacterial species. RESULTS All species of Streptococcus generally increased from ages 3 to 24 months. The relative ratios of the bacteria remained fairly constant at all ages studied (P > 0.1). There were no significant differences in numbers of all Streptococcus species between pre-term children and full-term controls at all the ages investigated between. CONCLUSION The results show that the relative numbers of S. mutans, S. sobrinus, S. mitis, S. sanguinis, and S. salivarius remain relatively constant from 3 to 24 months of age in caries-free pre- and full-term children.

Interleukin-1 and interleukin-1 inhibitor production by human adherent cells stimulated with periodontopathic bacteria

This study examined the effect of the putative periodontopathic bacteria Bacteroides gingivalis and Fusobacterium nucleatum on the production of interleukin-1 (IL-1) and IL-1 inhibitors by human plastic-adherent mononuclear cells from normal donors. Fusobacterium mortiferum was used as a non-oral, non-pathogenic control organism. Unstimulated adherent cells spontaneously secreted an IL-1 inhibitor, whereas stimulation with B. gingivalis induced the synthesis and secretion of IL-1. With both fusobacteria IL-1 was present in the intracellular environment, whereas the predominant secretory product was either IL-1 or an IL-1 inhibitor. These results suggest that bacteria are capable of modulating cytokine production by monocytes and may thereby alter the local immune response.

A microbiological study of Papillon-LeFevre syndrome in two patients

Aim—To analyse the microflora of subgingival plaque from patients with Papillon-Lefévre syndrome (PLS), which is a very rare disease characterised by palmar-plantar hyperkeratosis with precocious periodontal destruction. Methods—Bacterial isolates were identified using a combination of commercial identification kits, traditional laboratory tests, and gas liquid chromatography. Some isolates were also subjected to partial 16S rDNA sequencing. Plaque samples were also assayed for the presence of Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinomycetemcomitans in a quantitative enzyme linked immunosorbent assay (ELISA) using monoclonal antibodies. Results—The culture results showed that most isolates were capnophilic and facultatively anaerobic species—mainly Capnocytophaga spp and Streptococcus spp. The latter included S constellatus, S oralis, and S sanguis. Other facultative bacteria belonged to the genera gemella, kingella, leuconostoc, and stomatococcus. The aerobic bacteria isolated were species of neisseria and bacillus. Anaerobic species included Prevotella intermedia, P melaninogenica, and P nigrescens, as well as Peptostreptococcus spp. ELISA detected P gingivalis in one patient in all sites sampled, whereas A actinomycetemcomitans was detected in only one site from the other patient. Prevotella intermedia was present in low numbers. Conclusions—Patients with PLS have a very complex subgingival flora including recognised periodontal pathogens. However, no particular periodontopathogen is invariably associated with PLS.