Dentures are a Reservoir for Respiratory Pathogens.

PURPOSE: Recent studies have established a relationship between dental plaque and pulmonary infection, particularly in elderly individuals. Given that approximately one in five adults in the UK currently wears a denture, there remains a gap in our understanding of the direct implications of denture plaque on systemic health. The aim of this study was to undertake a comprehensive evaluation of putative respiratory pathogens residing upon dentures using a targeted quantitative molecular approach. MATERIALS AND METHODS: One hundred and thirty patients' dentures were sonicated to remove denture plaque biofilm from the surface. DNA was extracted from the samples and was assessed for the presence of respiratory pathogens by quantitative polymerase chain reaction (qPCR). Ct values were then used to approximate the number of corresponding colony forming equivalents (CFEs) based on standard curves. RESULTS: Of the dentures, 64.6% were colonized by known respiratory pathogens. Six species were identified: Streptococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, Haemophilus influenzae B, Streptococcus pyogenes, and Moraxella catarrhalis. P. aeruginosa was the most abundant species followed by S. pneumoniae and S. aureus in terms of average CFE and overall proportion of denture plaque. Of the participants, 37% suffered from denture stomatitis; however, there were no significant differences in the prevalence of respiratory pathogens on dentures between healthy and inflamed mouths. CONCLUSIONS: Our findings indicate that dentures can act as a reservoir for potential respiratory pathogens in the oral cavity, thus increasing the theoretical risk of developing aspiration pneumonia. Implementation of routine denture hygiene practices could help to reduce the risk of respiratory infection among the elderly population.

The Francis Report--Why it Matters to the Dental Team.

The Francis Report into the deaths at Mid-Staffordshire NHS Trust highlighted the problems facing the NHS when patients, families, clinicians and nurses are not heard, and where the management, leadership and ensuing culture are focused on the system's business, not patient care. This paper, the first in a series based on the implications of the Francis Report, provides the background and context for the subsequent 'perspective' articles from a range of relevant stakeholders and care-providers. The overriding message is that in all areas of healthcare, dentistry included, the quality of patient care, especially patient safety, must be placed above all other aims. Clinical Relevance: The overriding importance of patient-centredness and quality of care, above all other aims, is the key message of clinical relevance from the Francis Report.

In vitro Candida albicans biofilm induced proteinase activity and SAP8 expression correlates with in vivo denture stomatitis severity.

Denture stomatitis is a common inflammatory disorder of the palatal mucosa amongst denture wearers. The pathological changes are induced by Candida albicans biofilm on the fitting surface of the upper denture, and different individuals experience different levels of disease. C. albicans is known to produce secreted aspartyl proteinases (SAPs) to aid adhesion, invasion and tissue destruction. We hypothesised that differential expression and activity of SAPs from denture stomatitis isolates results in different levels of disease amongst denture wearers. We selected C. albicans isolates from asymptomatic controls and three different severities of disease [Newton's type (NT) 0, I, II and III]. We assessed biofilm formation and proteinase activity for each biofilm and investigated the transcriptional profile of SAPs 1, 2, 5, 6 and 8 from early (12 h) and mature (24 h) biofilms. There were no significant differences between isolates with respect to biofilm formation, whereas proteinase activity normalised to biofilm growth was significantly increased in the diseased groups (p < 0.0001). Proteinase activity correlated strongly with SAP expression (p < 0.0001). SAP8 expression was the greatest, followed by SAP5, 6, 2 and 1. The diseased groups showed the greatest levels of SAP expression, with significant differences also observed between the groups (p < 0.005). All SAPs except SAP5 were expressed in greater amounts in the mature biofilms compared to early biofilms. Overall, this study suggests that SAP activity in biofilms determined in vitro may help to explain differences in disease severity. SAP8 has been shown for the first time to play a prominent role in biofilms.

Molecular detection of transcriptionally active bacteria from failed prosthetic hip joints removed during revision arthroplasty.

The purpose of this study was to use microbiological culture and bacterial 16S rRNA gene sequencing methods to detect transcriptionally active bacteria present on the surface of failed prosthetic hip joints removed during revision arthroplasty. Five failed prosthetic hip joints were sonicated to dislodge adherent bacteria and subjected to microbiological culture. Bacterial RNA was extracted from each sonicate, cDNA prepared by reverse transcription and the 16S rRNA gene amplified using universal primers. Polymerase chain reaction (PCR) products were cloned, assigned to distinct groups by restriction fragment length polymorphism (RFLP) analysis and one representative clone from each group was sequenced. Bacteria were identified by comparison of the obtained 16S rRNA gene sequences with those deposited in public access sequence databases. All five specimens were positive for the presence of bacteria by both culture and PCR. Culture methods identified species from eight genera. Molecular detection of transcriptionally active bacteria identified a wider range of species. A total of 42 phylotypes were identified, of which Lysobacter gummosus was the most abundant (31.6%). Thirty-four clones (14.5%) represented uncultivable phylotypes. No potentially novel species were identified. It is concluded that a diverse range of transcriptionally active bacterial species are present within biofilms on the surface of failed prosthetic hip joints.

Reducing the incidence of denture stomatitis: are denture cleansers sufficient?

PURPOSE: Candida albicans is the predominant oral yeast associated with denture stomatitis. With an increasing population of denture wearers, the incidence of denture stomatitis is increasing. Effective management of these patients will alleviate the morbidity associated with this disease. The aim of this study was to examine the capacity of four denture cleansers to efficiently decontaminate and sterilize surfaces covered by C. albicans biofilms. MATERIALS AND METHODS: Sixteen C. albicans strains isolated from denture stomatitis patients and strain ATCC 90028 were grown as mature confluent biofilms on a 96-well format and immersed in Dentural, Medical Interporous, Steradent Active Plus, and Boots Smile denture cleansers according to the manufacturers' instructions or overnight. The metabolic activity and biomass of the biofilms were then quantified, and scanning electron microscopy (SEM) used to examine treated biofilms. RESULTS: Dentural was the most effective denture cleanser, reducing the biomass by greater than 90% after 20 minutes. Steradent Active plus was significantly more effective following 10-minute immersion than overnight (p < 0.001). All cleansers reduced the metabolic activity by greater than 80% following overnight immersion; however, Boots Smile exhibited significantly reduced metabolic activity following only a 15-minute immersion (p < 0.001). SEM revealed residual C. albicans material following Dentural treatment. CONCLUSIONS: This study showed that denture cleansers exhibit effective anti-C. albicans biofilm activity, both in terms of removal and disinfection; however, residual biofilm retention that could lead to regrowth and denture colonization was observed. Therefore, alternative mechanical disruptive methods are required to enhance biofilm removal.

Impact of frequency of denture cleaning on microbial and clinical parameters - a bench to chairside approach.

Objective: Robust scientific and clinical evidence of how to appropriately manage denture plaque is lacking. This two-part study (i) developed an in vitro model of denture plaque removal, and (ii) assessed effectiveness of these approaches in a randomised clinical trial. Method: (i) a complex denture plaque model was developed using the dominant microbial genera from a recent microbiome analyses. Biofilms formed on polymethylmethacrylate were brushed daily with a wet toothbrush, then either treated daily for 5 days or only on Days 1 and 5 with Polident® denture cleanser tablets (3 min soaking). Quantitative and qualitative microbiological assessments were performed. (ii), an examiner-blind, randomised, crossover study of complete maxillary denture wearers was performed (n = 19). Either once-daily for 7 days or on Day 7 only, participants soaked dentures for 15 min using Corega® denture cleansing tables, then brushed. Denture plaque microbiological assessment used sterilized filter paper discs. Results: The in vitro model showed daily cleaning with denture cleanser plus brushing significantly reduced microbial numbers compared to intermittent denture cleaning with daily brushing (p < 0.001). The clinical component of the study showed a statistically significant reduction in denture plaque microbial numbers in favour of daily versus weekly treatment (aerobic bacteria p = 0.0144). Both in vitro and in vivo studies showed that denture plaque biofilm composition were affected by different treatment arms. Conclusions: This study demonstrated that daily denture cleansing regimens are superior to intermittent denture cleansing, and that cleansing regimens can induce denture plaque compositional changes. Clinicaltrials.gov registration: NCT02780661.

Evaluating Streptococcus mutans Strain Dependent Characteristics in a Polymicrobial Biofilm Community.

Aim: The purpose of this study was to investigate strain dependent differences of the cariogenic biofilm forming Streptococcus mutans within both simple and complex communities. Methods: A mono-species containing representative S. mutans clinical isolates (caries and non-caries), and a multispecies in vitro caries biofilm model containing Lactobacillus casei, Veillonella dispar, Fusobacterium nucleatum and Actinomyces naeslundii, and either of two representative S. mutans clinical isolates (caries and non-caries), was developed as a comparison model. Compositional analysis of total and live bacteria within biofilms, and transcriptional analysis of biofilm associated virulence factors were evaluated by live/dead PCR and quantitative PCR, respectively. Scanning electron microscopy (SEM) was used to analyze the architecture of biofilm. One-way analysis of variance and t-tests were used to investigate significant differences between independent groups of data. Results: Within a mono-species biofilm, different S. mutans strains responded similarly to one another during biofilm formation in different carbohydrate sources, with sucrose showing the highest levels of biofilm biomass and galactose showing the lowest. Within the polymicrobial biofilm system, compositional analysis of the bacteria within the biofilm showed that S. mutans derived from a caries-free patient was preferentially composed of both total and viable L. casei, whereas S. mutans derived from a caries patient was dominated by both total and viable S. mutans (p < 0.001). Normalized gene expression analysis of srtA, gtfB, ftf, spaP, gbpB, and luxS, showed a general upregulation within the S. mutans dominant biofilm. Conclusion: We were able to demonstrate that individual strains derived from different patients exhibited altered biofilm characteristics, which were not obvious within a simple mono-species biofilm model. Influencing the environmental conditions changed the composition and functionality S. mutans within the polymicrobial biofilm. The biofilm model described herein provides a novel and reproducible method of assessing the impact on the biofilm microbiome upon different environmental influences.

Risk factors for chest infection in acute stroke: a prospective cohort study.

BACKGROUND AND PURPOSE: Pneumonia is a major cause of morbidity and mortality after stroke. We aimed to determine key characteristics that would allow prediction of those patients who are at highest risk for poststroke pneumonia. METHODS: We studied a series of consecutive patients with acute stroke who were admitted to hospital. Detailed evaluation included the modified National Institutes of Health Stroke Scale; the Abbreviated Mental Test; and measures of swallow, respiratory, and oral health status. Pneumonia was diagnosed by set criteria. Patients were followed up at 3 months after stroke. RESULTS: We studied 412 patients, 391 (94.9%) with ischemic stroke and 21 (5.1%) with hemorrhagic stroke; 78 (18.9%) met the study criteria for pneumonia. Subjects who developed pneumonia were older (mean+/-SD age, 75.9+/-11.4 vs 64.9+/-13.9 years), had higher modified National Institutes of Health Stroke Scale scores, a history of chronic obstructive pulmonary disease, lower Abbreviated Mental Test scores, and a higher oral cavity score, and a greater proportion tested positive for bacterial cultures from oral swabs. In binary logistic-regression analysis, independent predictors (P<0.05) of pneumonia were age >65 years, dysarthria or no speech due to aphasia, a modified Rankin Scale score > or =4, an Abbreviated Mental Test score <8, and failure on the water swallow test. The presence of 2 or more of these risk factors carried 90.9% sensitivity and 75.6% specificity for the development of pneumonia. CONCLUSIONS: Pneumonia after stroke is associated with older age, dysarthria/no speech due to aphasia, severity of poststroke disability, cognitive impairment, and an abnormal water swallow test result. Simple assessment of these variables could be used to identify patients at high risk of developing pneumonia after stroke.

The seroprevalence of hepatitis C virus infection among children and their mothers attending for dental care in Glasgow, Scotland, United Kingdom.

This paper describes a voluntary anonymous survey to investigate the seroprevalence of Hepatitis C (HCV) in children in Glasgow, UK attending a Dental Hospital and the proportion of HCV positive mothers who have a child who is HCV seropositive. The study was undertaken among children and accompanying parents and household contacts attending a general anaesthetic assessment clinic at Glasgow Dental Hospital and School. Children were asked to provide an oral fluid specimen for HCV testing. Accompanying adults were asked to provide demographic data on the child and information on familial risk factors for HCV infection using a standardised questionnaire. Birth mothers were also asked to provide an oral fluid specimen. Specimens and questionnaires were linked by a unique anonymous study number. Between June 2009 and December 2011, samples were collected from 2141 children and 1698 mothers. None of the samples from the children were HCV seropositive but 16 (0.9%, 95% CI 0.6-1.5%) of the specimens from mothers were HCV antibody positive. In summary, the prevalence of HCV seropositivity in the birth mothers of the children was similar to that estimated in the general population served by the hospital and showed no evidence of mother-to-child transmission of HCV.

Susceptibility to Melaleuca alternifolia (tea tree) oil of yeasts isolated from the mouths of patients with advanced cancer.

Yeasts that are resistant to azole antifungal drugs are increasingly isolated from the mouths of cancer patients suffering from oral fungal infections. Tea tree oil is an agent possessing antimicrobial properties that may prove useful in the prevention and management of infections caused by these organisms. In this study, 301 yeasts isolated from the mouths of 199 patients suffering from advanced cancer were examined by an in vitro agar dilution assay for susceptibility to tea tree oil. All of the isolates tested were susceptible, including 41 that were known to be resistant to both fluconazole and itraconazole. Clinical studies of tea tree oil as an agent for the prevention and treatment of oral fungal infections in immunocompromised patients merit consideration.

CJD: update for dental staff.

UNLABELLED: It is almost a decade since the recognition of the emergence of a new infectious disease termed variant Creutzfeldt-Jakob disease (vCJD) caused by prions (PrPTSE), abnormal variants of a normal human cell surface protein (PrP). This disease has a number of similarities to other forms of CJD--lethal disorders characterized by a prolonged incubation period, and progressive mental deterioration. In relation to oral tissues, PrPTSE have been found in neural, gingival, pulpal, lingual, lymphoreticular and salivary gland tissue in animal models. In both sporadic and variant CJD, PrPTSE is detectable in the trigeminal ganglion and, in vCJD, in lymphoreticular tissues, but infectivity has not been tested in other human oral tissues. CLINICAL RELEVANCE: PrPTSE is much more resistant to the common methods of inactivation than conventional pathogens, and it adheres avidly to steel whilst retaining its infectivity. Particular attention must be paid to cleaning and sterilizing re-usable dental instruments. Single-use devices, such as endodontic files and matrix bands, must never be re-used. Advice on the reprocessing of dental instruments used on known CJD patients must be obtained from local infection control teams. Research into effective methods of prion inactivation appears promising, although further work on the applicability to general dental practice is required.

Human Papilloma Virus (HPV) Oral Prevalence in Scotland (HOPSCOTCH): A Feasibility Study in Dental Settings.

The purpose of this study was to test the feasibility of undertaking a full population investigation into the prevalence, incidence, and persistence of oral Human Papilloma Virus (HPV) in Scotland via dental settings. Male and female patients aged 16-69 years were recruited by Research Nurses in 3 primary care and dental outreach teaching centres and 2 General Dental Practices (GDPs), and by Dental Care Teams in 2 further GDPs. Participants completed a questionnaire (via an online tablet computer or paper) with socioeconomic, lifestyle, and sexual history items; and were followed up at 6-months for further questionnaire through appointment or post/online. Saline oral gargle/rinse samples, collected at baseline and follow-up, were subject to molecular HPV genotyping centrally. 1213 dental patients were approached and 402 individuals consented (participation rate 33.1%). 390 completed the baseline questionnaire and 380 provided a baseline oral specimen. Follow-up rate was 61.6% at 6 months. While recruitment was no different in Research Nurse vs Dental Care Team models the Nurse model ensured more rapid recruitment. There were relatively few missing responses in the questionnaire and high levels of disclosure of risk behaviours (99% answered some of the sexual history questions). Data linkage of participant data to routine health records including HPV vaccination data was successful with 99.1% matching. Oral rinse/gargle sample collection and subsequent HPV testing was feasible. Preliminary analyses found over 95% of samples to be valid for molecular HPV detection prevalence of oral HPV infection of 5.5% (95%CI 3.7, 8.3). It is feasible to recruit and follow-up dental patients largely representative / reflective of the wider population, suggesting it would be possible to undertake a study to investigate the prevalence, incidence, and determinants of oral HPV infection in dental settings.

Voriconazole susceptibility of yeasts isolated from the mouths of patients with advanced cancer.

The in vitro activity of voriconazole was compared with those of fluconazole and itraconazole against 270 clinical isolates of yeasts from the mouths of patients receiving palliative care for advanced cancer. A broth micro-dilution assay as described by the National Committee for Clinical Laboratory Standards was employed for determination of MICs. Of the 270 isolates, 206 (76 %) were fluconazole sensitive and 64 were fluconazole resistant. Voriconazole showed more potent activity than either fluconazole or itraconazole, including against some isolates resistant to both fluconazole and itraconazole. However, for fluconazole-resistant isolates, the MICs of itraconazole and voriconazole were proportionally higher than for the fluconazole-susceptible isolates, suggesting cross-resistance. Voriconazole may be a useful additional agent for the management of oral fungal infections caused by strains resistant to fluconazole and itraconazole, but susceptibility cannot be assumed and in vitro MIC determination is recommended prior to its use.

Effectiveness of health care worker training on the oral health of elderly residents of nursing homes.

OBJECTIVES: The aim of this study was to evaluate the effect of a staff training programme on mouth care on the oral health of elderly residents of long-term care institutions. METHODS: Seventy-eight residents of five long-stay institutions were enrolled and underwent a baseline oral health assessment. Staff caring for residents in three of the sites received intensive training in mouth care. This comprised lecture and video material complemented by clinical demonstrations. The oral health of residents at all five sites was reassessed at periods of 3 and 9 months. Staff caring for residents in the remaining two institutions were then provided with mouth care training and all patients were reassessed at 18 months. Statistical analyses were undertaken to examine for significant changes in selected oral health parameters after training, within each group. RESULTS: Oral mucosal disease and oral dryness were common at baseline. The staff training was well received. Following staff training, there was a significant reduction in the number of residents left to undertake their own oral care. There were significant improvements in denture hygiene and a reduction in the number of residents wearing dentures overnight. The prevalence of oral mucosal disease dropped, with significant reductions in angular cheilitis and denture stomatitis. CONCLUSION: This education programme was effective in changing oral health care procedures within long-stay institutions for the elderly, with measurable improvements in oral health of the residents.

Training in oral disease, diagnosis and treatment for medical students and doctors in the United Kingdom.

To find out if the training of medical undergraduates and qualified doctors was adequate to diagnose, investigate, manage, and refer common oral disorders appropriately, we sent anonymous questionnaires to undergraduate and postgraduate medical and dental deans, accident and emergency (A&E) doctors, and dentists. We wanted to know if they were capable of diagnosing and treating 10 common oral disorders, and if their training was adequate to enable them to do so. Ten clinical photographs with short clinical histories were sent to 48 A&E physicians together with a structured questionnaire. Twenty-one of the 29 medical schools in the UK responded to a questionnaire about the teaching given in the current curriculum about oral anatomy and pathology, and the prevention of oral disease. A questionnaire sent to the deans of the 16 British dental schools asked how many academic staff were involved in undergraduate teaching, and how many in postgraduate courses. A third questionnaire was sent to the 24 postgraduate medical deans to find out how many postgraduate courses there were for qualified medical staff. Of the 48 medical staff, 134 (28%) diagnosed cases correctly, compared with 194 (88.7%) of the 22 dentists, indicating serious deficiencies in diagnostic awareness. Only 11 of the 21 medical schools who responded currently incorporate teaching of oral pathology in their curricula. We conclude that doctors and medical students are inadequately educated about oral diseases with obvious consequences.

Evaluation of the recurrence of denture stomatitis and Candida colonization in a small group of patients who received itraconazole.

OBJECTIVES: The aim of the study was to determine the recurrence rate of denture stomatitis and persistence of Candida in 22 patients (5 male and 17 female, mean age 71 years) over a 3-year period. STUDY DESIGN: Denture hygiene practice, denture cleanliness, and the presence of palatal erythema were assessed for each patient at the start of the study (baseline). The oral cavity was sampled for yeasts by imprint culture and denture discs. Ten patients received a capsular form of itraconazole (100 mg twice daily for 15 days) and 12 patients were provided with 100 mg of itraconazole in the form of a mouthwash (10 mL twice daily), which was then swallowed. No further antifungal treatment was administered to any of the patients. Clinical and microbiological assessments were repeated for each patient at 6 months and 3 years after the original appointment. Yeasts were identified by colony color on CHROMagar Candida, germ-tube formation, and API-32C profiling. Selected isolates were then typed by inter-repeat polymerase chain reaction (IR PCR). RESULTS: Candida albicans was isolated at baseline from all patients either alone (12 patients) or in combination with another species (10 patients). Other yeast species recovered were C glabrata (5 patients), C tropicalis (1 patient), C guilliermondii (1 patient), C krusei (1 patient), C parapsilosis (1 patient), C kefyr (1 patient), and Saccharomyces cerevisiae (2 patients). Candida albicans and/or C glabrata were recovered from 11 of the 22 patients after 6 months or 3 years. A complete and consistent change of yeast species from baseline was observed in 6 patients after 6 months and at 3 years. The remaining 5 patients were yeast-free at the follow-up assessments. PCR fingerprinting of C albicans and C glabrata indicated strain persistence over 6 months in 10 patients and in 4 patients after 3 years. A switch in strain type occurred for 1 patient after 6 months and for 3 patients after 3 years. CONCLUSIONS: The recurrence of denture stomatitis in patients who maintained a high standard of denture cleanliness was low. Although itraconazole was beneficial in reducing the fungal load, there may be strain persistence or subsequent recolonization of the oral cavity by a broader range of potentially less sensitive yeast species.

Prions and the human transmissible spongiform encephalopathies.

The last 5 years have seen the emergence of a new disease in humans (vCJD), mainly in the United Kingdom. This emergence has been accompanied by an explosion of scientific data on a novel group of the responsible infectious agents called prions and has profound implications for infection control and transfusion policies. Also of concern is the finding of prions in neural, gingival, pulpal, and salivary tissue in animal models and significant titers of infectivity from extraneural organs (particularly, in cases of vCJD, in lymphoreticular tissues). There is limited information on the presence of prion proteins in the oral tissues from human studies. Because of the differences in patterns of disease in animal models and in strains of prion protein, it is difficult to extrapolate directly these findings to humans, but it illustrates a potential for transmission by way of the dental route. High levels of infectivity may be present in tissues early in the incubation period and before clinical signs and symptoms. The dental profession must turn its attention to the routine decontamination of dental instruments to ensure that these procedures are performed to the highest regulatory standard. Clinicians and manufacturers must work closely together to develop instruments that are either single use or can be presented in a form that can be more easily decontaminated. Clinicians must pay close attention to manufacturers' decontamination instructions and must not reuse items designated as single use, such as endodontic files. Improvements in compliance with these requirements will not only reduce the risk of transmission of TSEs but also other less tenacious infectious agents.

CJD and the dentist.

The last 10 years have seen the emergence of a new disease termed variant CJD. This disease is thought to be initiated by abnormal prion proteins. This article reviews the different clinical manifestations of human prion diseases and provides some background information on the biological nature of this unique infectious agent. Prion proteins present a challenge to infection control because of their relative resistance to the conventional sterilization process. This article highlights the issues involved and discusses current guidelines for management of patients with human transmissible encephalopathies.