The evolving new understanding of endodontic infections

The science of oral microbiology is in a period of change from the era of bacterial cultivation to an era of molecular genetic methods and techniques. Already a significant body of new knowledge exists with regard to the oral flora in health and disease. Inevitably, this new knowledge has led to a better understanding of many oral diseases. In endodontics, the prevailing concepts are still to a great extent based on the results of the classical cultivation studies. However, a few groups have started to use molecular methods, and a new understanding of endodontic infections is presently evolving. Thus, the root canal infection clearly is more complex than revealed by cultivation methods alone, and both previously unidentified and uncultivable microorganisms have been detected by molecular methods. A reasonable estimate at present is that the infected root canal contains, not less than 10, but rather between 10 and 50 bacterial species which coincide well with the number of bacterial species normally found in a dental plaque sample and at different sites in the oral cavity. A further interesting finding in the studies using molecular techniques is that the microbiota of the infected root canal appears to be very similar to the flora of the periodontal pocket in patients with active periodontal disease. With regard to infection of periapical lesions in patients with asymptomatic apical periodontitis, electron microscopic and molecular methods have confirmed our cultivation findings that this is a common occurrence. Mature biofilms have been demonstrated on the external surfaces of root tips and in the form of sulfur granules within periapical granulomas. As in dental plaque, Actinomyces species appear to have a special role as scaffold builders in the development of sulfur granules. Other bacteria are then attracted to the site and a multibacterial granule (biofilm) develops. In addition, in situ hybridization studies show a variety of different bacteria and bacterial morphotypes in periapical lesions. With DNA–DNA hybridization between 11and 39 bacterial species have been recognized in the lesions, again confirming that in patients with active disease, the microbiotas of endodontic and periodontal infections are very similar. Thus, the recent findings demonstrate and confirm that the periapical endodontic lesion is not as hostile to microorganisms as many have thought. As clinicians we have to understand and accept that an infection might not be limited to the root of the tooth, but include the periapical lesion as well.     

Polymerase chain reaction‐based denaturing gradient gel electrophoresis in the evaluation of oral microbiota

Background/aims: Clinical evaluation of oral microbial reduction after a standard prophylactic treatment has traditionally been based on bacterial cultivation methods. However, not all microbes in saliva or dental plaque can be cultivated. Polymerase chain reaction‐based denaturing gradient gel electrophoresis (PCR‐DGGE) is a cultivation‐independent molecular fingerprinting technique that allows the assessment of the predominant bacterial species present in the oral cavity. This study sought to evaluate the oral microbial changes that occurred after a standard prophylactic treatment with a conventional oral care product using PCR‐DGGE. Methods: Twelve healthy adults participated in the study. Pooled plaque samples were collected at baseline, 24 h after prophylaxis (T1), and 4 days after toothbrushing with fluoride toothpaste (T4). The total microbial genomic DNA of the plaque was isolated. PCR was performed with a set of universal bacterial 16S rDNA primers. The PCR‐amplified 16S rDNA fragments were separated by DGGE. The effects of the treatment and of dental brushing were assessed by comparing the PCR‐DGGE fingerprinting profiles. Results: The mean numbers of detected PCR amplicons were 22.3 ± 6.1 for the baseline group, 13.0 ± 3.1 for the T1 group, and 13.5 ± 4.3 for the T4 group; the differences among the three groups were statistically significant (P < 0.01). The study also found a significant difference in the mean similarities of microbial profiles between the baseline and the treatment groups (P < 0.001). Conclusion: PCR‐based DGGE has been shown to be an excellent means of rapidly and accurately assessing oral microbial changes in this clinical study.     

Technology Development to Explore the Relationship Between Oral Health and the Oral Microbial Community

The human oral cavity contains a complex microbial community that, until recently, has not been well characterized. Studies using molecular tools have begun to enumerate and quantify the species residing in various niches of the oral cavity; yet, virtually every study has revealed additional new species, and little is known about the structural dynamics of the oral microbial community or how it changes with disease. Current estimates of bacterial diversity in the oral cavity range up to 700 species, although in any single individual this number is much lower. Oral microbes are responsible for common chronic diseases and are suggested to be sentinels of systemic human diseases. Microarrays are now being used to study oral microbiota in a systematic and robust manner. Although this technology is still relatively young, improvements have been made in all aspects of the technology, including advances that provide better discrimination between perfect-match hybridizations from non-specific (and closely-related) hybridizations. This review addresses a core technology using gel-based microarrays and the initial integration of this technology into a single device needed for system-wide studies of complex microbial community structure and for the development of oral diagnostic devices.     

Population studies of microbial ecology in periodontal health and disease

It has been established that the bacterial diversity in any given environment is severely underestimated when assessed by means of culture-based techniques. Yet, almost all currently available knowledge related to the periodontal microbiota in health and disease has been generated either by culture-based surveys or by methods that require prior species identification by culture. A handful of recent studies using culture-independent molecular methods providing 16S rRNA sequences for both cultivable and not yet cultivated species of human periodontal bacteria demonstrated a high bacterial diversity in the oral cavity. It has been estimated that approximately 500 species may colonize the human oral cavity, half of which have been cultivated to date. A review of the available epidemiological data on the prevalence of certain periodontal microbiota on a population level reveals considerable variation in estimates with respect to 1) sampling strategy, 2) mode of bacterial identification, and 3) race/ethnicity of the studied population. Nevertheless, specific bacterial profiles appear to confer high odds ratios for pathological periodontal conditions and/or progressive periodontal disease. However, the currently recognized periodontal pathogens are commonly recovered from periodontally healthy children, and their carrier rate in adults is substantial. Virulent clones, such as a highly leukotoxic strain of Actinobacillus actinomycetemcomitans, have been found to be closely associated with aggressive forms of periodontitis. In conclusion, while the majority of the periodontal microbiota are commensals, a subset of likely opportunistic pathogens fulfills the epidemiologic requirements needed in order to be ascribed as risk/causative factors. Given the large proportion of the periodontal microbial habitat that is currently insufficiently explored, and assuming that the hitherto uncultivated segment of the bacterial community will include similar levels of pathogenic species, the list of periodontal pathogens should be expected to expand.     

Using DGGE profiling to develop a novel culture medium suitable for oral microbial communities

More than 700 bacterial species have been detected in the human oral cavity. They form highly organized microbial communities and are responsible for many oral infectious diseases, such as dental caries and periodontal disease. The prevention and treatment of these diseases require a comprehensive knowledge of oral microbial communities, which largely relies on culture-dependent methods to provide detailed phenotypic and physiological analysis of these communities. However, most of the currently available laboratory media can only selectively support the growth of a limited number of bacterial species within these communities, and fail to sustain the original oral microbial diversity. In this study, using denaturing gradient gel electrophoresis (DGGE) as an index to systematically survey and analyse the selectivity of commonly used laboratory media, we developed a new medium (SHI medium) by combining the ingredients of several selected media that can support different subpopulations within the original oral microbial community derived from pooled saliva. DGGE and 454 pyrosequencing analysis showed that SHI medium was capable of supporting a more diversified community with a microbial profile closer to that of the original oral microbiota. Furthermore, 454 pyrosequencing revealed that SHI medium supported the growth of many oral species that have not before been cultured. Crystal violet assay and the confocal laser scanning microscope analysis indicated that, compared with other media, SHI medium is able to support a more complex saliva-derived biofilm with higher biomass yield and more diverse species. This DGGE-guided method could also be used to develop novel media for other complex microbial communities.     

The effect of a microbead dentifrice on microbial load in oral microenvironments

To cite this article:
Int J Dent Hygiene 9 , 2011; 136–142
DOI: 10.1111/j.1601‐5037.2010.00465.x
Sreenivasan PK, Haraszthy VI, Zambon JJ.The effect of a microbead dentifrice on microbial load in oral microenvironments. Abstract: Objectives: The human oral cavity contains several microenvironments or ecologic niches. While mechanical plaque control is well known to reduce the number of supragingival dental plaque bacteria, there is little data on antimicrobial effects in other oral ecologic niches. The present study examined the effects of mechanical plaque control using a microbead dentifrice on bacteria colonizing oral ecologic niches. Methods: Twenty‐two adults (aged 18–70 years) including nine generalized moderate chronic periodontitis subjects and 13 periodontally healthy subjects having average gingival indices ≥1 and plaque indices ≥1.5 completed a 1 week washout phase and refrained from oral hygiene the morning of baseline sample collection. Microbial samples from supragingival dental plaque, buccal mucosa, dorsal surface of the tongue and whole mixed saliva were obtained. Subjects brushed with a microbead dentifrice and, after 10 min, sampling was repeated. The number of anaerobic bacteria was determined by culture on non‐selective media and transformed to log₁₀ for statistical analyses. Results: Mechanical plaque control using the microbead dentifrice resulted in statistically significant reductions in bacterial numbers in each ecologic niche (P < 0.001). The greatest reduction in the number of viable bacteria occurred in samples taken from the buccal mucosa (97.22%) followed by a 95.22% reduction in supragingival plaque bacteria, a 94.51% reduction in the number of bacteria on the dorsal surface of the tongue and a 91.57% reduction in the number of bacteria in whole mixed saliva. Conclusions: Mechanical plaque control using a microbead dentifrice reduces microbial load in microenvironments throughout the human oral cavity.     

Does assessment of microbial composition of plaque/saliva allow for diagnosis of disease activity of individuals?

Abstract Microbiological tests are limited in their applicability in the assessment of caries activity and in caries prediction. They can be effective in groups of persons with high or low caries experience. The reasons for the limitation of microbiological tests rests with unique characteristics of the microflora and local environments of the oral cavity, which will modify the cariogenicity of plaque in an individual. Thus, high numbers of S. mutans may be associated with the development of a lesion at a site, while a second susceptible site with high levels of this organism in the same subject will remain caries free. This paper identifies some aspects of oral bacteria which can contribute to the unique nature of the microflora associated with plaque in an individual. Firstly, the range of bacteria potentially involved in caries has widened and now includes, for example, ‘low pH’ non-mutans streptococci. The presence of such organisms in plaque in an individual may influence early enamel demineralization. Most significantly. Streptococcus mutans, Streptococcus mitis and Actinomyces naeslundii have been shown to be comprise many distinct clones, with different distribution among subjects. Little is known of the impact of clonal diversity on caries activity but in some bacterial diseases particular clones are associated with virulence. Therefore, possession of a particular clone or clones by an individual could be related to caries activity. Also, the extent of clonal diversity may reflect the nature of the oral environment. Recent studies suggest that cells are released from biofilms during adherence and growth, i.e. the early phases of development. Thus, determination of the numbers of a given species in non-stimulated saliva may indicate whether it is actively growing in plaque. Microbiological tests on the oral flora should perhaps be used to monitor the status of the oral cavity, after establishing a norm for the individual patient. Research on species and clonal diversity of oral bacteria among human populations; diversity and its role in the caries process; and the liberation of biofilm cells could provide data to allow better appreciation and evaluation of the results of microbiologiocal testing.     

变性梯度凝胶电泳分析口腔微生物多样性的研究进展

变性梯度凝胶电泳(DGGE)是近些年微生物分子生态学研究中的热点技术之一.由于DGGE技术具有可靠性强、重现性高和方便快捷等优点,被广泛地应用于微生物群落多样性和动态性分析.下面就DGGE的原理、在口腔微生物研究中的应用、优缺点和应用前景作一.     

Invasion of dentinal tubules by root canal bacteria

Bacterial invasion of dentinal tubules and the clinical consequences have been recognized for over a century. However, while many components of the infected dentinal tubule microflora have been identified, it is likely that there are etiological agents involved in endodontic infections that have not yet been recognized. Bacterial invasion of coronal dentinal tubules occurs when the dentine is exposed to the oral environment and of radicular dentinal tubules subsequent to infection of the root canal system or as a consequence of periodontal disease. The content and architecture of a dentinal tubule can influence bacterial invasion, with tubule patency being important. This can account for regional variations in bacterial invasion and is particularly seen with dentinal sclerosis, where more advanced sclerotic changes in apical radicular tubules, especially in elderly individuals, limit bacterial invasion in this area. While several hundred bacterial species are known to inhabit the oral cavity, a relatively small and select group of bacteria are involved in invasion of dentinal tubules. Gram‐positive organisms dominate the tubule microflora in both carious and non‐carious dentine. The relatively high numbers of obligate anaerobes present, such as Eubacterium spp., Propionibacterium spp., Bifidobacterium spp., Peptostreptococcus micros, and Veillonella spp., suggests that the environment favors the growth of these bacteria. Gram‐negative obligate anaerobic rods, e.g. Porphyromonas spp., are less frequently recovered; however, with time, fastidious obligately anaerobic bacteria become established as principal components of the microflora and can be found within the deep dentine layers. In the early stages of infection, Gram‐positive bacteria dominate the microflora. The identification of adhesins that mediate these initial interactions of bacteria with dentine is important for understanding the development of tubule infection and in designing adhesion‐blocking compounds. Recent evidence suggests that streptococci and enterococci may recognize components present within dentinal tubules, such as collagen type I, which stimulate bacterial adhesion and intra‐tubular growth. Specific interactions of other oral bacteria with invading streptococci may then facilitate invasion of dentine by select bacteria. It is important therefore that the mechanisms of invasion and inter‐bacterial adhesion are understood to assist development of novel control strategies.     

Isolation of C. dubliniensis from insulin-using diabetes mellitus patients

The identification of the novel candidal species, C. dubliniensis, from oral swab studies of HIV-seropositive and -seronegative individuals has led to speculation that such a species may also reside in the oral cavity of other patient groups. In this study involvement of the newly described species, C. dubliniensis, was investigated in oral carriage and disease in 414 insulin-using diabetes mellitus patients. Seventy-seven percent of the diabetic patients carried candidal species in the oral cavity. C. albicans was the most commonly identified candidal species. This was followed by C. dubliniensis, which was isolated on 64 occasions. Colonisation with multiple candidal species was common, and C. dubliniensis was present in both carriage and disease states. Seven patients without signs of oral disease had C. dubliniensis isolated as the sole candidal species, while the same species was associated with various forms of oral candidosis in six patients.     

Polymerase chain reaction-based denaturing gradient gel electrophoresis in the evaluation of oral microbiota

BACKGROUND/AIMS: Clinical evaluation of oral microbial reduction after a standard prophylactic treatment has traditionally been based on bacterial cultivation methods. However, not all microbes in saliva or dental plaque can be cultivated. Polymerase chain reaction-based denaturing gradient gel electrophoresis (PCR-DGGE) is a cultivation-independent molecular fingerprinting technique that allows the assessment of the predominant bacterial species present in the oral cavity. This study sought to evaluate the oral microbial changes that occurred after a standard prophylactic treatment with a conventional oral care product using PCR-DGGE. METHODS: Twelve healthy adults participated in the study. Pooled plaque samples were collected at baseline, 24 h after prophylaxis (T1), and 4 days after toothbrushing with fluoride toothpaste (T4). The total microbial genomic DNA of the plaque was isolated. PCR was performed with a set of universal bacterial 16S rDNA primers. The PCR-amplified 16S rDNA fragments were separated by DGGE. The effects of the treatment and of dental brushing were assessed by comparing the PCR-DGGE fingerprinting profiles. RESULTS: The mean numbers of detected PCR amplicons were 22.3 +/- 6.1 for the baseline group, 13.0 +/- 3.1 for the T1 group, and 13.5 +/- 4.3 for the T4 group; the differences among the three groups were statistically significant (P < 0.01). The study also found a significant difference in the mean similarities of microbial profiles between the baseline and the treatment groups (P < 0.001). CONCLUSION: PCR-based DGGE has been shown to be an excellent means of rapidly and accurately assessing oral microbial changes in this clinical study.     

Identification of Candida species in the clinical laboratory: a review of conventional,commercial, and molecular techniques

In healthy individuals, Candida species are considered commensal yeasts of the oral cavity. However, these microorganisms can also act as opportunist pathogens, particularly the so‐called non‐albicans Candida species that are increasingly recognized as important agents of human infection. Several surveys have documented increased rates of C. glabrata, C. tropicalis, C. guilliermondii, C. dubliniensis, C. parapsilosis, and C. krusei in local and systemic fungal infections. Some of these species are resistant to antifungal agents. Consequently, rapid and correct identification of species can play an important role in the management of candidiasis. Conventional methods for identification of Candida species are based on morphological and physiological attributes. However, accurate identification of all isolates from clinical samples is often complex and time‐consuming. Hence, several manual and automated rapid commercial systems for identifying these organisms have been developed, some of which may have significant sensitivity issues. To overcome these limitations, newer molecular typing techniques have been developed that allow accurate and rapid identification of Candida species. This study reviewed the current state of identification methods for yeasts, particularly Candida species.     

Chronic Periodontal Disease,Periodontal Pathogen Colonization,and Increased Risk of Precancerous Gastric Lesions

Background: This study assesses the association between periodontal pathogen colonization and the potential risk of developing precancerous lesions of gastric cancer (PLGC) in a clinical setting. Methods: Included were 35 newly diagnosed patients with PLGC and 70 age‐matched individuals without PLGC. A full‐mouth intraoral examination was performed to assess periodontal conditions. Stimulated whole saliva and pooled plaque samples were collected to evaluate colonization by Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans and to characterize oral microbial diversity in saliva and dental plaque. Results: Compared with the control group, patients with PLGC experienced higher prevalence of bleeding on probing (31.5% versus 22.4%; P <0.05), higher levels of T. denticola (P <0.01) and A. actinomycetemcomitans (P <0.01), and less bacterial diversity in their saliva (P <0.01). The final multivariate logistic regression model consisting of all key sociodemographic characteristics, oral health behavioral factors, and periodontal assessments revealed that elevated colonization with periodontal pathogens, specifically T. forsythia, T. denticola, and A. actinomycetemcomitans, decreased bacterial diversity in dental plaque, and not flossing teeth regularly was a significant predictor of increased risk of PLGC (P = 0.022). Conclusion: Findings of the present study provide new evidence suggesting that periodontal pathogen burdens and bacterial diversity in the oral cavity are important factors contributing to a potentially increased risk of developing precancerous gastric lesions.     

Streptococcus mutans copper chaperone,CopZ, is critical for biofilm formation and competitiveness

The oral cavity is a dynamic environment characterized by hundreds of bacterial species, saliva, and an influx of nutrients and metal ions such as copper. Although there is a physiologic level of copper in the saliva, the oral cavity is often challenged with an influx of copper ions. At high concentrations copper is toxic and must therefore be strictly regulated by pathogens for them to persist and cause disease. The cariogenic pathogen Streptococcus mutans manages excess copper using the copYAZ operon that encodes a negative DNA‐binding repressor (CopY), the P1‐ATPase copper exporter (CopA), and the copper chaperone (CopZ). These hypothetical roles of the copYAZ operon in regulation and copper transport to receptors led us to investigate their contribution to S. mutans virulence. Mutants defective in the copper chaperone CopZ, but not CopY or CopA, were impaired in biofilm formation and competitiveness against commensal streptococci. Characterization of the CopZ mutant biofilm revealed a decreased secretion of glucosyltransferases and reduced expression of mutacin genes. These data suggest that the function of copZ on biofilm and competitiveness is independent of copper resistance and CopZ is a global regulator for biofilm and other virulence factors. Further characterization of CopZ may lead to the identification of new biofilm pathways.     

Antibiotic effects on bacterial profile in osteonecrosis of the jaw

Oral Diseases (2011) 18 , 85–95 Objective: Oral infection is considered to play a critical role in the pathogenesis of bisphosphonate‐related osteonecrosis of the jaw (BRONJ), and antibiotic therapy has become a mainstay of BRONJ therapy. This study was aimed to investigate the effect of antibiotics on bacterial diversity in BRONJ tissues. Materials and methods: The bacterial profile from soft tissues associated with the BRONJ lesion was determined using 16S rRNA‐based denaturing gradient gel electrophoresis (DGGE) and sequencing. Twenty BRONJ subjects classified as stage 0–2 were enrolled in this study, and patient groups were divided into an antibiotic cohort (n = 10) treated with systemic antibiotic and a non‐antibiotic cohort (n = 10) with no prior antibiotic therapy. Results: The DGGE fingerprints indicated no significant differences in bacterial diversity of BRONJ tissue samples. Patients on antibiotics had higher relative abundance of phylum Firmicutes with bacterial species, Streptococcus intermedius, Lactobacillus gasseri, Mogibacterium timidum, and Solobacterium moorei, whereas patients without antibiotics had greater amounts of Parvimonas micra and Streptococcus anginosus. Thirty percent of bacterial populations were uncultured (yet‐to be cultured) phylotypes. Conclusion: This study using limited sample size indicated that oral antibiotic therapy may have a limited efficacy on the bacterial population associated with BRONJ lesions.     

Molecular profiling of oral microbiota in jawbone samples of bisphosphonate-related osteonecrosis of the jaw

Oral Diseases (2012) 18, 602–612 Objective: Infection has been hypothesized as a contributing factor to bisphosphonate (BP)‐related osteonecrosis of the jaw (BRONJ). The objective of this study was to determine the bacterial colonization of jawbone and identify the bacterial phylotypes associated with BRONJ. Materials and methods: Culture‐independent 16S rRNA gene‐based molecular techniques were used to determine and compare the total bacterial diversity in bone samples collected from 12 patients with cancer (six, BRONJ with history of BP; six, controls without BRONJ, no history of BP but have infection). Results: Denaturing gradient gel electrophoresis profile and Dice coefficient displayed a statistically significant clustering of profiles, indicating different bacterial population in BRONJ subjects and control. The top three genera ranked among the BRONJ group were Streptococcus (29%), Eubacterium (9%), and Pseudoramibacter (8%), while in the control group were Parvimonas (17%), Streptococcus (15%), and Fusobacterium (15%). H&E sections of BRONJ bone revealed layers of bacteria along the surfaces and often are packed into the scalloped edges of the bone. Conclusion: This study using limited sample size indicated that the jawbone associated with BRONJ was heavily colonized by specific oral bacteria and there were apparent differences between the microbiota of BRONJ and controls.     

Clinical relevance of surface characteristics on the formation of plaque on teeth and implants

The mouth is, from an ecologic point of view, an 'open growth system' with a continuous transport of micro-organisms. To cause infection (caries or periodontitis) or even to survive in the oral cavity, micro-organisms need to attach to one of the available surfaces, otherwise they will be removed by a continuous flow of saliva. The mouth, with his ideal growth conditions for micro-organisms, has many places, called niches, that can be colonised with micro-organisms. It is not surprising that the mouth is being colonised with more then 400 different species. This article discusses successively colonisation of the oral cavity, bacterial adhesion, plaque growth from a clinical point of view, the influence of surface properties on the initial plaque adhesion and maturation, and finally important factors influencing the plaque formation on oral implants.     

Molecular analysis of human oral microbiota

OBJECTIVES: The application of molecular, mainly 16S ribosomal RNA (rRNA)-based approaches enables researchers to bypass the cultivation step and has proven its usefulness in studying the microbial composition in a variety of ecosystems, including the human oral cavity. In this mini-review, we describe the impact of these culture-independent approaches on our knowledge of the ecology of the human oral cavity and provide directions for future studies that should emphasize the role of specific strains, species and groups of microbes in periodontal disease. MATERIALS AND METHODS: Recent findings are summarized to elucidate the relationship between periodontal disease and human oral microbiota, including as-yet-to-be-cultured organisms. RESULTS: The real-time polymerase chain reaction (PCR) method was developed to detect and quantify periodontopathic bacteria, such as Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis (formerly Bacteroides forsythus) and Treponema denticola. The checkerboard DNA-DNA hybridization technique allowed enumeration of large numbers of species in very large numbers of samples. 16S rRNA gene clone library analysis revealed the diversity of human oral microbiota and the existence of as-yet-to-be-cultured organisms that are presumed periodontal pathogens. In addition, terminal restriction fragment length polymorphism (T-RFLP) analysis was applied for assessment of diversity of human oral microbiota. CONCLUSION: Culture-independent approaches are useful for studying the microbial ecology in the human oral cavity and should be useful in the future to elucidate the etiology of periodontal disease.     

Clonal persistence of oral Fusobacterium nucleatum in infancy

Once established, early-colonizing bacterial species tend to persist in the mouth. To obtain detailed information on the population dynamics of early-colonizing oral anaerobes, we examined the clonal diversity and persistence of clones among oral Fusobacterium nucleatum populations during the first 2 yrs of life. Consecutive salivary samples from 12 infants, collected at 2, 6, 12, 18, and 24 mos of age, yielded a total of 546 F. nucleatum isolates for clonal typing with arbitrarily primed PCR (AP-PCR). Up to 7 AP-PCR types were simultaneously detected in each sample. In 11 out of the 12 infants examined, AP-PCR types persisted for up to 1 yr. Strain turnover rate was high during the first year of life, but then the occurrence of persistent clones increased. This study indicates a wide genetic diversity within the species and provides evidence for the increasing persistence of F. nucleatum clones in the oral cavity with age.     

Isolation and identification of Candida from the oral cavity

A number of methods of sampling the oral cavity for the presence of candida have been developed. Such techniques play an important role in the diagnosis and management of oral candidosis. In the past, identification of candida isolated from the oral cavity has usually been limited to the genus Candida or to the species C. albicans. However, with the recognition that Candida species differ in the production of putative virulence factors and sensitivity to antifungal agents, greater emphasis has been placed on identification of isolates to species level. As a result a range of commercially available systems for yeast identification can now be used in conjunction with traditional identification procedures.