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Oral microbiology: current concepts in the microbiology of dental caries and periodontal disease.

The microbial flora of the mouth is highly complex, containing a wide variety of bacterial species. The most common types of oral disease, dental caries and periodontal disease, are both related to dental plaque and seem to occur when the normal balance between the microorganisms and the host is disturbed in some way. Dental caries is usually associated with increased numbers of mutans streptococci and lactobacilli at the sites of disease; estimation of salivary levels of these organisms may be useful for assessing caries risk in patients and for monitoring their response to preventive measures. A large number of 'candidate pathogens' have been identified as potential aetiological agents in different types of periodontal disease, although the 'specific plaque hypothesis' may still be controversial. Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, together with the poorly understood spirochaetes, have most frequently been reported as significant periodontopathogens and a number of possible virulence factors have been described. Application of modern molecular techniques to the study of the microbiology of oral diseases should allow rapid further progress to be made and will lead, hopefully, to improved methods of diagnosis, risk assessment and treatment.

Animals↗

Oral microorganisms and cardiovascular disease.

The list of etiological factors for cardiovascular disease is long, complicated, intertwined, and yet to be completed. This paper will evaluate the current evidence for the pathogenic role of certain microorganisms, including those of the oral cavity, in the etiology of cardiovascular disease.

Bacterial Infections↗

Micropunctures of rubber gloves used in oral surgery.

In 720 operations in ambulatory oral surgery, the 2,880 surgical rubber gloves used were tested at the end of operation for perforations not earlier detected (microperforations). Gloves of ordinary thickness (type A) and of a thicker quality (type B) were examined. Perforations were found in 1495% and 22.0% of type A gloves and in 3.2% and 2.0% of type B gloves when oral surgeons and dental students, respectively, were operators. Gloves worn by operation assistants showed a lower incidence of perforation. Before operation, 50 gloves of type A were punctured at the tip of right index finger by a sterile dental probe. Results of bacteriologic examinations of the right index and middle fingers strongly indicated that a high number of bacteria passed through pinholes in the gloves. The intact surgical glove represents an impermeable barrier, protecting not only the operation wound against skin bacteria from the hands of the oral surgeon, but also the oral surgeon against pathogenic microorganisms, in particular hepatitis virus type B, escaping from the oral cavity of the patient.

Antisepsis↗

Biology of oral mucosa and esophagus.

The mucosal lining of the oral cavity and esophagus functions to protect the underlying tissue from mechanical damage and from the entry of microorganisms and toxic materials that may be present in the oropharynx. In different regions, the mucosa shows adaptation to differing mechanical demands: Masticatory mucosa consists of a stratified squamous keratinized epithelium tightly attached to the underlying tissues by a collagenous connective tissue, whereas lining mucosa comprises a nonkeratinized epithelium supported by a more elastic and flexible connective tissue. The epithelium is constantly replaced by cell division in the deeper layers, and turnover is faster in the lining than in the masticatory regions. Chemotherapeutic agents and radiation limit proliferation of the epithelium so that it becomes thin or ulcerated; this will first occur in the lining regions. The principal patterns of epithelial differentiation are represented by keratinization and nonkeratinization. As keratinocytes enter into differentiation, they become larger and begin to flatten and to accumulate cytokeratin filaments. In addition to the keratins, the differentiating keratinocytes synthesize and retain a number of specific proteins, including profilaggrin, involucrin, and other precursors of the thickening of the cell envelope in the most superficial layers. The concept of epithelial homeostasis implies that cell production in the deeper layers will be balanced by loss of cells from the surface. There is a rapid clearance of surface cells, which acts as a protective mechanism by limiting colonization and invasion of microorganisms adherent to the mucosal surface.

Antineoplastic Agents↗

Efficacy of ozone on survival and permeability of oral microorganisms.

In the present study, we examined the effect of ozonated water on oral microorganisms and dental plaque. Almost no microorganisms were detected after being treated with ozonated water (4 mg/l) for 10 s. To estimate the ozonated water-treated Streptococcus mutans, bacterial cells were stained with LIVE/DEAD BacLight Bacterial Viability Kit. Fluorescence microscopic analysis revealed that S. mutans cells were killed instantaneously in ozonated water. Some breakage of ozonated water-treated S. mutans was found by electron microscopy. When the experimental dental plaque was exposed to ozonated water, the number of viable S. mutans remarkably decreased. Ozonated water strongly inhibited the accumulation of experimental dental plaque in vitro. After the dental plaque samples from human subjects were exposed to ozonated water in vitro, almost no viable bacterial cells were detected. These results suggest that ozonated water should be useful in reducing the infections caused by oral microorganisms in dental plaque.

Aggregatibacter actinomycetemcomitans↗

Simonsiella filaments isolated from erosive lesions of the human oral cavity.

The morphology and cultural characteristics of bacteria of the Simonsiella genus isolated from erosions of oral mucosa are reported here. Direct microscopic examination of smeared oral swabs and the consequent selection of suitable culturing procedures and media are compulsory for recovering Simonsiella microorganisms from clinical specimens in routine work.

Bacterial Infections↗

Effect of delmopinol on in vitro dental plaque formation, bacterial acid production and the number of microorganisms in human saliva.

This study investigated the effect of a surface active compound, delmopinol, on plaque formation and established plaque in vitro, on the bacterial acid production from glucose and on the total viable bacterial counts in saliva. The antimicrobial effect was compared with that of chlorhexidine. The plaque-inhibiting effect was evaluated in an artificial mouth system, and the effect on bacterial acid production was registered as a decrease of pH in bacterial suspensions with various concentrations of delmopinol. It was shown that delmopinol is able to prevent plaque formation, to dissolve established plaque in vitro, and that it has 5-125 times higher minimum inhibitory concentrations than chlorhexidine. Saliva samples collected 1 min after rinsing with delmopinol showed on differences in the total number of bacteria in saliva as compared with controls. It was also shown that the bacterial acid production from glucose was reduced successively with increasing concentrations of delmopinol. The results indicate that delmopinol might be as effective as chlorhexidine against plaque formation and that delmopinol is capable of penetrating established plaque, thus promoting a more effective mechanical cleansing.

Anti-Infective Agents, Local↗

Bacterial diversity in aortic aneurysms determined by 16S ribosomal RNA gene analysis.

BACKGROUND: Aortic aneurysms are common vascular conditions that cause considerable morbidity and mortality. Understanding of the mechanisms involved in the pathogenesis of the condition remains limited. Recently, infection has been suggested as possible contributor in the development of the disease. The aim of the present study was to examine aortic aneurysms for the presence of bacterial DNA using polymerase chain reaction (PCR) targeting the 16S ribosomal RNA (rRNA) gene, followed by cloning and sequencing. METHODS: Universal eubacterial primers were used to amplify 16S rRNA bacterial genes in 10 specimens from arterial walls of aortic aneurysms. Subsequently, PCR amplicons were cloned into Escherichia coli and sequencing of the cloned inserts was used to determine species identity or closest relatives by comparison with known sequences in GenBank. RESULTS: Sequences of Stenotrophomonas spp., including S. maltophilia (formerly Pseudomonas homology group V) were detected in six aneurysm samples. Propionibacterium acnes was identified in five samples, and Brevundimonas diminuta (formerly P. diminuta) in four samples. Other species previously assigned to the Pseudomonas genus such as Comamonas testosteroni, Delftia acidovorans, Burkholderia cepacia, Herbaspirillum sp., and Acidovorax sp. were also detected. Some clones fell into other environmental species, including Methylobacterium sp. and Bradyrhizobium elkanii, and others represented bacteria that have not yet been cultivated. DNA sequences from oral bacteria, including Streptococcus sanguinis, Tannerella forsythia, and Leptotrichia buccalis were detected. Sequences from Prevotella melaninogenica and Lactobacillus delbrueckii, which are commonly found in both mouth and gastrointestinal tract, were also detected. Additional species included Dermacoccus spp. and Corynebacterium vitaeruminis. CONCLUSIONS: A wide variety of bacteria, including oral bacteria, was found to colonize aortic aneurysms and may play a role in their development. Several of these microorganisms have not yet been cultivated. CLINICAL RELEVANCE: Although Chlamydophila pneumoniae has been detected in aneurysmal walls, its exact role in the condition remains inconclusive. Overall, there is scarce information about the role of microorganisms in aneurysmal disease. In the present study, we used molecular genetics to detect a diversity of bacteria in arterial walls of aortic aneurysms. The presence of multiple microorganisms in aneurysmal disease may have implications for chemoprophylaxis and antibiotic treatment if directed only at C.pneumoniae.

Adult↗

Outbreak of cryptosporidiosis linked with a farm event.

AIM: To investigate an outbreak of cryptosporidiosis linked to a two-day farm educational event in the Wellington region. Methods. The outbreak was investigated by carrying out a site visit and interviewing cases (or their parents) identified through notifications to Wellington Regional Public Health. RESULTS: Twenty confirmed cases were linked to the event. Nineteen were aged under seven years. The most likely route of infection was the hand-to-mouth transfer of the parasite after touching an infected animal. CONCLUSIONS: To our knowledge this is the first outbreak of cryptosporidiosis associated with a farm event reported in New Zealand. Farm animals, particularly calves, are likely to carry cryptosporidiosis and other microorganisms that are pathogenic in humans. This outbreak highlights the need for organisers of similar events to implement preventive measures such as hand washing facilities and reminders. Adults supervising young children should also be vigilant in enforcing hand washing following animal contact.

Adolescent↗

Possible association between amniotic fluid micro-organism infection and microflora in the mouth.

OBJECTIVE: To determine whether oral bacteria are found in the amniotic cavity. DESIGN: Laboratory based analysis of clinical samples. SETTING: Royal London Hospital, Whitechapel. POPULATION: Forty-eight women attending for elective caesarean section. METHODS: Dental plaque, a high vaginal swab, amniotic fluid and chorioamnion tissue were taken from women with intact membranes. MAIN OUTCOME MEASURES: Samples were investigated using culture and microscopy for the presence of microorganisms. Amniotic fluid was analysed by polymerase chain reaction (PCR) for the presence of the ubiquitous 16S rRNA gene specific to most eubacteria. Samples were analysed using PCR genus and species specific primers directed to bacterial taxa found as part of the normal oral microflora (Streptococcus spp. and Fusobacterium nucleatum). Levels of prostaglandin E2 and cytokines were measured in amniotic fluid. RESULTS: Amniotic fluid was positive for universal bacteria PCR, Streptococcus spp. PCR and F. nucleatum PCR in 34/48, 20/48 and 7/48 of cases, respectively. Streptococcus spp. and F. nucleatum were cultured from the dental plaque, vagina and amniotic fluid of 48/48, 14/48, 0/48 and 29/48, 6/48, 0/48 subjects, respectively. A significant association was found between detection of microbial DNA (universal and F. nucletum) and complications in previous pregnancies including miscarriage, intrauterine death, neonatal death, preterm delivery and premature rupture of membranes (P < 0.05 and P < 0.01, respectively). Prostaglandin E2 and cytokine levels, with the exception of IL-1alpha, were not significantly different between women with and without evidence of infection. CONCLUSIONS: The results indicate that Streptococcus spp. and F. nucleatum in the amniotic fluid may have an oral origin.

Adult↗

A retrospective investigation of advanced periodontal disease as a risk factor for septicemia in hematopoietic stem cell and bone marrow transplant recipients.

OBJECTIVE: Septicemia is a cause of death in hematopoietic stem cell transplant (HSCT) recipients. Extraction of teeth with advanced periodontitis has been advocated before HSCT to prevent septicemia in myeloablated hosts. The primary aim of the present study was to determine impact of chronic periodontitis, as measured by radiographic alveolar bone loss, on septicemia and transplant mortality. STUDY DESIGN: A retrospective design was used to study 77 subjects who received pretransplant dental evaluation, panoramic radiography, and full myeloablative allogeneic HSCT to treat hematologic malignancies. Radiographic crestal alveolar bone loss was measured with a Schei ruler on all teeth. Microorganisms isolated from positive blood cultures within the first 100 days after transplant were categorized as of likely origin from periodontal, oral, or any body sites. Spearman correlation and logistic regression analysis assessed associations between positive blood cultures, mean subject whole-mouth percent radiographic crestal alveolar bone loss, and 100-day survival. RESULTS: Radiographic crestal alveolar bone loss per study subject averaged 13% +/- 7%, with 18.2% exhibiting bone loss of 20% or greater. During the initial 100 days after transplant, 63.6% subjects yielded septicemia-associated positive blood cultures, with Staphylococcus epidermidis, Streptococcus mitis, Enterococcus faecalis, Streptococcus sanguis, Staphylococcus aureus, and Escherichia coli as the most common isolates recovered. No statistically significant associations were found between mean subject radiographic alveolar bone loss and septicemia of likely periodontal or oral origin. CONCLUSION: In this preliminary study, no relationship was found between radiographic periodontal status and septicemia or mortality within the initial 100 days after transplant. A larger-sized, prospective study is warranted to further delineate the risk of septicemia from periodontal and other oral diseases in immunocompromised patients.

Adult↗

Angular cheilitis: a clinical and microbial study.

The purpose of this prospective study was to re-examine the relative importance of various factors in the pathogenesis of angular cheilitis. Sixty-four patients with cheilitis were examined clinically and microbiologically. In addition, a subsample of 23 patients was examined for serum iron and transferrin. The clinical appearance of the lip lesions fell into 4 categories. A ground rhagad at the corner of the mouth involving adjacent skin, was the most frequent type among dentate patients, whereas among denture wearers a deep lesion following the labial marginal sulcus was frequently observed. Dentate patients and denture wearers with cheilitis often had atopic constitution or cutaneous diseases. Pathogenic microorganisms were cultured from the lesions in all 64 patients; Staphylococcus aureus in 40 patients and Candida albicans in 45. The results of this study indicate a correlation between angular cheilitis and pathogenic microorganisms. Furthermore, among dentate patients, a correlation exists between cutaneous discomfort and angular cheilitis. Other etiological factors suggested for this disorder were found to be of subordinate importance.

Adolescent↗

Prevalence, distribution of serotypes, and cariogenic potential in hamsters of mutans streptococci from elderly individuals.

The prevalence of mutans streptococci (Streptococcus mutans, Streptococcus cricetus, Streptococcus sobrinus, and Streptococcus rattus) was determined in the salivas of 169 elderly individuals ranging in age from 60 to 87 years. Approximately 40% of these individuals were edentulous and wore full upper and lower dentures. With the exception of a higher proportion of saliva counts below 1,000 CFU/ml in the full-denture wearers, the prevalence and the serotype and species distributions of the mutans streptococci were similar in the denture wearers and individuals with natural teeth only. The species and serotype distributions of mutans streptococci in this elderly population were also consistent with reported observations of other workers on younger, more caries-prone populations. A total of 87 representative isolates of the mutans streptococci were tested for cariogenic potential in a hamster model system. A considerable degree of variation in virulence between different strains was observed. However, these differences were not relatable to individual species or serotypes or to whether the organisms were isolated from denture wearers or naturally dentate subjects. The results of our studies indicate that elderly individuals with either natural or artificial dentitions may be a hitherto unrecognized reservoir of mutans streptococci having varying degrees of potential cariogenicity. Hence, in close family situations they could serve, along with parents and siblings, as vectors in the initial transmission of cariogenic microorganisms to young children.

Aged↗

Susceptibility of viridans streptococci of 27 antimicrobial agents.

One hundred and eighty strains of viridans Streptococci (Str. mitis, Str. salivarius, Str. MG, Str. SBE, Str. bovis, Str. equinus and Str. uberis) were isolated from mouth and upper respiratory tract secretions of humans at random in different parts of Poland. These strains were tested for susceptibility to 27 antimicrobial agents. Analysis of results was performed with special reference to bacterial endocarditis, which is caused mostly by these microorganisms. At low minimal inhibitory concentrations, most effective antibiotics appeared amoxycillin, clindamycin, azidocillin, cephaloridine, ampicillin, erythromycin and penicillin G. Susceptibility to other penicillins and cephalosporins was much less pronounced. Small part of strains was highly susceptible to tetracyclines, whereas all other antimicrobial agents were practically ineffective.

Anti-Bacterial Agents↗

Longitudinal study of prognostic factors in established periodontitis patients.

Numerous indicators for disease progression have been described in the last decade. The purpose of this study was to examine, longitudinally, a large battery of clinical, microbiological, and immunological indicators, to try to determine whether the presence of one or a combination of these parameters at baseline, would correlate positively with increased attachment and or bone loss (true prognostic factors). Following initial screening, 79 patients with established periodontitis were monitored longitudinally for one year. Whole mouth clinical measurements, plaque gingival and calculus indices, together with pocket depth and attachment level measurements, were repeated every three months. Full mouth radiographic survey, performed at baseline and 12 months, served to determine changes in crestal bone height using an image enhancement technique. Subgingival plaque samples were taken at baseline and every 3 months. Immunofluorescence assays were performed for the a battery of target microorganisms. Serum and GCF samples for IgG subclasses analysis were obtained at each visit and assayed using ELISA techniques. Likewise blood, samples were also drawn at each visit for a quantitative analysis of serum cotinine level. The overall mean attachment loss (AL) and bone loss (BL) were almost identical (0.159 mm and 0.164 mm, respectively). Individual patients variation was large (-0.733 to +1.004 mm). An overall 6.89% of sites were active; individual patients' means ranged from 0-28.9%. Mean pocket depth (PD) showed minimal change over the study period (-0.033 mm) thus suggesting that most if not all the AL was accompained by concomitant gingival recession. Smokers exhibited greater AL and radiographic BL compared to non-smokers. Likewise, patients' cotinine level showed direct correlation with outcomes of progressive periodontal breakdown. Past severity of periodontal involvement, as reflected in the patients baseline PD, AL and crestal bone height, showed good correlation with longitudinal changes in the periodontium. This correlation was higher for crestal BL as the outcome variable, while somewhat smaller for change in AL as the outcome variable. Bacteroides forsythus (Bf.), Prevotella intermedia (Pi.) and Porphyromonas gingivalis (Pg.) were frequently found in these patients. The presence of these microorganisms at baseline was associated with further disease progression. Subjects with mean baseline pocket depth equal or greater than 3.2 mm were at greater risk for future bone loss 1 year later (O.R. 2.97; C.I. 1.02-8.70). Smokers were at significantly greater risk for further attachment loss when compared to non-smokers (O.R. 5.41; C.I. 1.50-19.5). Subjects that harbored B. forsythus at baseline, were at seven times greater risk for increased pocket depth (O.R. 7.84; C.I.1.74-35.3). In conclusion, past periodontal destruction, smoking habits, Bf., Pg., & Pi. are prognostic factors for further periodontal breakdown. When designing clinical trials, or when evaluating epidemiological data, it is most important to balance for these factors. Also, treatment strategies should attempt to eliminate or modify these factors.

Adult↗