Microbiological impressions of teeth, saliva and dietary fibre can predict caries activity.

OBJECTIVES: Mutans streptococci and Lactobacilli on impressions of teeth and caries tests were used to identify caries prone subjects. METHODS: Twenty-seven dental students were examined for caries initially and after 4 years. At the initial examination plaque index; saliva flow, buffering capacity and lysozyme; sucrose and fibre consumption; Lactobacilli and mutans streptococci in saliva and on alginate impressions were measured. Data was analysed using Kruskal-Wallis, Wilcoxon-Mann-Whitney and Spearman's Rank correlation tests and linear discriminant analysis. RESULTS: The best predictor of caries increment was decay. Nine subjects had no caries or restored teeth with caries (Group A); 9 had restored teeth with no caries but developed an average of 8 new decayed surfaces (Group B); 9 had an average of 4.4 decayed surfaces and developed a further 9.6 (Group C). Group A had fewer filled surfaces than Group B (p=0.02) and Group C (p=0.024) a higher flow rate of stimulated saliva than Group B (p=0.02) and Group C (p=0.012). Microorganisms were cultured from all decayed teeth, 98% that developed decay, 89% filled and 69% sound teeth. Fibre intake, saliva flow and the percentage of teeth or sound teeth with Lactobacilli and mutans streptococci gave a specificity of 89%, a sensitivity of 100% and predicted an increase in decay in all Group B subjects while one subject from Group A was misclassified. When teeth with microorganisms were excluded four subjects were misclassified. CONCLUSIONS: Growth of cariogenic microorganisms on alginate impressions, saliva flow and dietary fibre predicted caries activity in most subjects.

(B1) Candida and mycotic infections.

Oral candidiasis (OC) is the most common mucosal manifestation of HIV infection. This workshop examined OC and other mycoses associated with HIV infection. Historically, blood CD4 cell numbers were the primary prognosticator for the development of OC. However, a study that statistically evaluated the predictive role of HIV viral load vs. CD4 cell counts revealed viral load to be a stronger predictor for OC. The role of biofilms and antifungal resistance in recalcitrant OC is unclear at present. In general, micro-organisms including yeasts in biofilms are more resistant to antifungals than their planktonic counterparts. When the remaining organisms are eliminated, the few resistant organisms may not be problematic, because they are present in low numbers. Unusual exotic mycoses in HIV-infected patients are more common in patients from the developing than the developed world. These infections may be recurrent and recalcitrant to therapy, be present in multiple and uncommon sites, increase with the progression of HIV disease, and may play a role similar to that of the more common mycoses. Typing and subtyping of yeasts are probably not critical to the clinical management of candidiasis caused by Candida albicans and non-albicans strains, including C. dubliniensis, because it is responsive to antifungal therapy. C. glabrata is probably the only exception. The presence of oral thrush in infants younger than 6 months of age is associated with an increased post-natal transmission risk of HIV infection. Thus, perinatal retroviral therapy should be combined with the treatment of oral thrush to prevent the post-natal acquisition of HIV.

Isolation of pathogenic Legionella species and legionella-laden amoebae in dental unit waterlines.

Legionella released into the air during treatment are a potential source of infection. Water stagnation in dental unit waterlines (DUWLs) creates biofilms and promotes the proliferation of these micro-organisms. This study investigated the presence of amoeba infected with legionella, L. pneumophila and other pathogenic Legionella species in a dental teaching hospital. Water samples were collected in the morning and afternoon from 99 dental units and 16 taps connected to the municipal water supply. Samples were plated on selective media and tested for legionella using the direct immunofluorescent antibody technique and the latex agglutination test. Legionella were found in 33% of the DUWLs and in 47% of the mains taps supplying these units. Legionella-laden amoebae occurred in one mains tap sample and in 20% of DUWLs in a clinic of the teaching hospital. L. micdadei was the predominant species isolated from this clinic. L. pneumophila serogroups 2-14 predominated in the mains water, whereas L. pneumophila serogroup 1 was found in approximately half of the contaminated DUWLs and mains taps irrespective of the time of sampling. Pathogenic Legionella species seeded by municipal water into DUWLs is a potential source of legionella infection for both dental personnel and patients during prolonged dental treatment. This problem is compounded by the presence of legionella-laden amoebae which may contain levels of organism well within the infective dose. The interaction of legionella with amoebae is an important ecological factor that may significantly increase the risk of legionellosis, and thus should be given further consideration in the refinement of risk assessment models.

Vomeronasal and olfactory nerves of adult and larval bullfrogs: I. Axons and the distribution of their glomeruli.

The distribution of primary olfactory and vomeronasal (VMN) glomeruli is demonstrated in the forebrain of the larval (stage III tadpole) and adult bullfrog. Olfactory glomeruli are massed at the anterior end of the olfactory bulb; farther posterior, they are located at the periphery and then become distributed in a superficial layer all along the ventral surface of the bulb. Glomeruli of the accessory olfactory bulb (AOB) of the tadpole are less compact and show larger dendritic profiles and fewer synaptic junctions per unit area than those of the adult AOB. Axons of both tadpole and adult VMN nerve are ultrastructurally indistinguishable from those of the primary olfactory nerve; in comparing VMN axons of tadpole and adult, however, differences were observed in the number of neurofilament profiles in axons at the proximal and distal ends of the VMN nerve. Counts of neurofilament profiles in the two ends of the nerve suggest that neurofilament segments accumulate near terminations of their axons. The VMN nerve of tadpole and adult frog are roughly the same size, and calculations indicate that they contain comparable numbers of axons. The primary olfactory nerve of the tadpole is estimated to contain about 660,000 axons; the adherent VMN nerve of the tadpole contains about 270,000 axons (a ratio of about 2.5:1). In the adult, the ratio of primary to VMN axons is about 64:1, reflecting the quantitative growth of the primary olfactory nerve with development from an aquatic tadpole to an air-breathing adult.

Structural studies of amyloglucan and a soluble glucan produced from starch by Streptococcus sanguis 1 MC 204.

Two alpha-D-glucans, produced from amylopectin by an oral isolate Streptococcus sanguis 1 MC 204, were shown to contain both (1----4) and (1----6) linkages. The first alpha-D-glucan (amyloglucan) was adherent and highly insoluble, and methylation analysis, i.r. spectroscopy, and enzymic analyses showed it to be similar to amylopectin but less branched with longer interior and exterior chains. The second polymer was a non-adherent soluble alpha-D-glucan that was similar to amyloglucan but with long exterior chains. These alpha-D-glucans were not synthesised de novo, but were the products of the modification of amylopectin.

Efficacy of three surface disinfectants for dental radiographic films and gloves.

Contaminated radiographic films and gloves may transmit infectious diseases. Objective. To determine whether Pre Sept (NaDC), a sodium dichloroisocyanurate based disinfectant; Bronocide SP (AIP) an alcohol-phenol-iodine disinfectant and polyvinyl pyrrolidine iodine (PVPI) can be used to disinfect radiographic films and gloves. Study design. Radiographic films were contaminated with Candida albicans, Streptococcus mutans, Lactobacilli or saliva and placed in either 0.1, 0.25 or 0.5 NaDC for 1, 2 or 5 min; PVPI for 5 min or sprayed with AIP. Gloves contaminated with C. albicans, S. mutans or Lactobacilli were sprayed with AIP. After treatment the films and gloves were tested for viable microorganisms. Results. A 0.5% solution of NaDC killed all microorganisms after one-minute exposure. PVPI killed 99.8% and AIP spray between 95.8 and 99% of microorganisms. Conclusions. NaDC was the most successful disinfectant in the laboratory and clinical setting followed by PVPI and AIP.

The effect of elastomeric impression materials on the growth of micro-organisms.

The influence of elastomeric impression materials on the growth of micro-organisms was examined in vitro. Bacillus subtilis was inoculated into broth containing impression materials and incubated at 37 degrees C for 72 hours. Express STD Putty, President Putty and Jet-Light Body, Low and Very High Viscosity Permagum and Provil L stimulated growth whereas Impregum-F and Express Light Body inhibited growth. The influence of Impregum-F and Express Light Body on oral micro-organisms was investigated further. Broth extracts were prepared by soaking these materials in Todd Hewitt broth for either 5 or 10 minutes. Thereafter, the extracts were inoculated with oral strains of Staphylococcus aureus, Streptococcus mutans and Candida albicans, incubated at 37 degrees C for 72 hours and plated on blood and Sabourauds agar to test for the presence of viable micro-organisms. The 10-minute broth extracts killed all the test isolates which suggests that impressions taken with Impregum-F and Express Light Body may not require disinfecting.

A note on a code for the identification of the 'viridans' streptococci.

A simple numerical code for the identification of 'viridans' streptococci was devised with Facklam's system of classification. This is based on biochemical criteria and consists of nine tests, with an additional five for the identification of closely related species and the subspecies or biotype of Streptococcus mutans. The 'viridans' streptococci can be divided into 10 species with the aid of this scheme while five subspecies and an additional biotype of Streptococcus mutans can be recognized. Advantages of this scheme are that the numerical code differentiates between isolates of the same species, distinguishes between biochemically active and non-active species and indicates similarities between species.

Antibacterial properties of eight dental cements.

The antibacterial action of a light-cured glass polyalkenoate Vitrebond and an adhesive resin luting agent Panavia Ex was investigated and compared with six previously tested dental cements. The bacterial cultures used were six strains of Streptococcus mutans, six of Streptococcus sanguis and six cultures of carious debris from carious lesions. A modification of the agar diffusion method was used to test the antibacterial action of freshly mixed cements. In addition the cements were allowed to set and their antimicrobial action was tested by incubation for 5 days in sucrose broth inoculated with the test cultures. All the freshly mixed materials had an antibacterial action. A two-way analysis of variance and Tukey-Student range analysis showed that Vitrebond had a significantly greater zone of inhibition than Aquacem, IRM, Dycal, Dycal VLC, Ceramco and GC Elite (P = 0.001). Freshly mixed Panavia Ex exhibited minimal antibacterial action. After 5 days the antibacterial properties of all the cements were reduced.

Yeast counts as a measure of host resistance in dental patients.

This study was undertaken to determine whether yeast counts could be used as an indicator of decreased host resistance. One hundred subjects were investigated, 20 young healthy dentate and 20 edentulous adults, 20 adult and 20 geriatric denture wearers and 20 patients fitted with obturators and dentures. Eleven of these obturators were silicone and 9 were acrylic. The denture and the palate of each subject was sampled by taking Sabourauds agar impressions of the posterolateral palatal area and incubating the impressions aerobically at 37 degrees C for 48 hours. The number of yeast colonies were counted and the findings analysed statistically. Palates and dentures of debilitated patients showed high yeast counts and it is concluded that a decreased host resistance can be determined by the method used in this investigation.

Salivary and plaque acids in caries active and caries free subjects.

The levels of organic acids in the plaque and saliva of 30 subjects aged between 12 and 20 years were investigated. Fifteen of the subjects were caries free and 15 had active carious lesions. The variables examined were the DMFT, plaque index, bleeding index, the flow rate of saliva, daily intake of fibre and sucrose and the level of acids in resting plaque and saliva exposed to a glucose rinse. Posterior bitewing radiographs were taken to confirm the presence of carious lesions. The Kruskal Wallis one-way analysis of variance was used to compare caries free and caries active subjects. Caries active subjects consumed more sucrose (p < 0.001) and had lower levels of acetic acid in their saliva (p = 0.05) than caries free subjects. The acetic acid-acetate buffer system can act as a buffer which suggests that acetic acid in the saliva of caries free subjects may diffuse into plaque to act as an effective buffer to counteract the destructive effects of lactic acid.

Serum and salivary antibodies to a mycobacterial 65-kDa stress protein are elevated in HIV-positive patients and modified by oral candidiasis.

Serum immunoglobulin G (IgG) and IgA, and salivary IgA antibodies to a mycobacterial stress protein (mSP65) were determined in human immunodeficiency virus (HIV)-positive patients, acquired immunodeficiency syndrome (AIDS) patients and HIV-negative controls with or without oral candidiasis. Serum IgG antibodies were elevated in patients with HIV infection and AIDS and especially in subjects with candidiasis compared with controls (P < 0.02, P < 0.005). This was not apparent with serum IgA. In the absence of candidiasis, salivary IgA antibodies were elevated in HIV-positive patients compared with AIDS (P < 0.005) patients and healthy controls (P = 0.001). The relative avidity of serum IgG antibodies to mSP65 in controls with candidiasis was lower than healthy controls (P < 0.0001). In saliva there was a decrease in the relative avidity of IgA antibodies in AIDS patients with candidiasis compared with HIV patients (P < 0.03). In patients without candidiasis, the relative avidity was higher in HIV patients than healthy controls (P = 0.02). The results suggest that HIV infection leads to raised serum and salivary antibodies to heat shock proteins. Concurrent Candida infection may modify both the titer and relative avidity differently for serum and saliva.

Immunoglobulin A (IgA), IgA1, and IgA2 antibodies to Candida albicans in whole and parotid saliva in human immunodeficiency virus infection and AIDS.

Human immunodeficiency virus (HIV)-infected individuals are predisposed to recurrent oral candidiasis, and, although it has been assumed that this is because of deficient mucosal immune responses, this has not been properly established. The present study aimed to compare the concentrations and secretion rates of immunoglobulin A (IgA) and IgA subclass antibodies to Candida albicans in whole and parotid saliva samples from HIV-infected patients, AIDS patients, and control subjects. Levels of IgA antibody to Candida species in whole saliva were higher in the HIV group than in the controls and were highest in the AIDS group (P < 0.05). In parotid saliva, the mean antibody levels were significantly greater in HIV-positive patients than in controls (P < 0.05) but fell to lower levels in the AIDS group. The secretion rates of Candida antibodies in parotid saliva were reduced in AIDS patients compared with HIV patients. The specific activities of the IgA antibodies and both subclasses were significantly higher in the HIV and AIDS patients than in the controls in both whole and parotid saliva (P < 0.05). Antibody levels were significantly correlated with the numbers of Candida organisms isolated from saliva (P < 0.05). These results suggest clear differences in salivary antibody profiles among HIV-infected. AIDS, and control subjects and are indicative of a response to antigenic challenge by infecting Candida species. No obvious defect in the mucosal immune response in the HIV or AIDS groups that might account for the increased prevalence of candidiasis was apparent.

Disinfection of dental diamond burs contaminated with hepatitis B virus.

STATEMENT OF PROBLEM: Hepatitis B infection (HBV) is a significant hazard in the dental environment because the virus may be transmitted through contaminated dental instruments. PURPOSE: This study determined whether cold disinfectants can inactivate HBV DNA and HBV surface antigens on diamond burs contaminated with HBV and whether ultrasonication can increase the antiviral properties of these agents. MATERIAL AND METHODS: Sterile dental diamond burs were contaminated with serum from a patient who tested positive for HBV surface antigen and hepatitis B viral DNA. The burs were air dried and placed in solutions containing either Cidex, Asepsys, TBS, Rotagerm, Virkon disinfectants, or a control phosphate buffered saline. Burs were divided into 2 groups and disinfected for 15 minutes. The first group was ultrasonicated; the second group was not ultrasonicated during disinfection. All the burs were transferred to phosphate buffered saline and ultrasonicated to remove any remaining viral particles. The ultrasonicate was tested for the presence of HBV surface antigen with a microparticle enzyme immunoassay and for hepatitis B viral DNA with a chemiluminescent molecular hybridization assay. RESULTS: TBS did not require ultrasonication to inactivate viral DNA and surface antigen. Rotagerm and Virkon inactivated surface antigen and viral DNA only with ultrasonication. Cidex and Asepsys inactivated viral DNA but not surface antigen with ultrasonication. CONCLUSION: The chlorine containing compound TBS was the most active disinfectant tested and did not require ultrasonication to destroy HBV. The remaining disinfectants should be used with ultrasonication to inactivate HBV.

The effect of titanium on the growth of plaque micro-organisms in vitro.

The effect of titanium on the growth of plaque organisms was studied in vitro. Thioglycollate broth containing either titanium or glass cylinders was inoculated with plaque micro-organisms and incubated aerobically and anaerobically at 37 degrees C for 6 hours. Growth was measured by observing an increase in turbidity of the broth. Cultures grew more vigorously aerobically than anaerobically. In the presence of titanium there was also more growth aerobically than anaerobically. There was no significant difference between the turbidity values obtained in the presence or absence of titanium suggesting that titanium does not influence the growth of plaque micro-organisms significantly.

Fungal infections associated with HIV infection.

Oral candidiasis is perhaps the commonest infection seen in HIV disease. The aim of this workshop was to provide a sketch of the multifarious aspects of the disease from a global perspective. To this end the panellists addressed issues such as the virulence of Candida, emergence of antifungal resistance, management of candidiasis and other exotic, oral mycotic diseases. An all-pervasive theme was the dramatic differences in the management of fungal infections consequential to the availability (or the lack) of anti-HIV drugs in the developed and the developing world. Further, the social stigmata associated with the HIV disease in many developing regions in Africa and Asia appears to modify the therapeutic strategies. Additionally, the lesser-known regional variations in the disease manifestations and therapeutic approaches were stark. Further work is direly needed to address these issues.