Prevalence and susceptibility profiles of oral yeast species isolated from a healthy adult Thai cohort.

OBJECTIVE: As the emerging resistance of Candida species to common antifungals is a major global concern, we assessed the antifungal susceptibility of oral yeast isolates from a healthy, Thai adult cohort, and correlated the yeast prevalence with oral disease indices. METHODS: Oral rinse samples collected from 100 Thai adults were concentrated and cultured on CHROMagar Candida. The yeasts were then isolated, identified and finally speciated using Matrix Assisted Laser Desorption ionization-time of flight mass spectrometry. Their antifungal sensitivity against fluconazole, itraconazole, voriconazole, and amphotericin B were investigated using standard Etest strips. The decayed, missing, filled teeth (DMFT) and the periodontal health were recorded and correlated with mycological data. RESULTS: The overall oral yeast prevalence was 25%. C. albicans was the commonest species isolated, followed by C. tropicalis and C. dubliniensis. Non-albicans-Candida was noted in approximately one-third, and included C. lusitaniae and C. nivariensis; Trichosporon asahii, was also detected in one subject. A majority of C. albicans isolates, (> 54%), exhibited resistance to fluconazole and voriconazole, while approximately a quarter (27%) were resistant to itraconazole. The vast majority (92%) however, were susceptible to amphotericin B. Those with oral yeasts had a significantly higher DMFT score (p < 0.05). CONCLUSION: The resistance of a majority of Candida spp. to common azoles, described here for the first time in a Thai cohort, is disconcerting, and appear to confirm the creeping emergence of antifungal resistance globally. An incidental finding was the positive correlation between oral yeast carriage and DMFT score in Thai subjects.

Profuse diversity and acidogenicity of the candida-biome of deep carious lesions of Severe Early Childhood Caries (S-ECC).

Introduction: The retentive niches of deep caries lesions have a distinct biome. Methods: We evaluated the site-specific (occlusal and proximal) Candida-biome of Severe-Early Childhood Caries (S-ECC) in 66- children (132 lesions). Asymptomatic primary molars fitting the definition of the International Caries Detection and Assessment-(ICDAS)-caries-code 5/6 were analyzed. Deep-dentinal sampling and simultaneous assessment of pH were performed. Clinical isolates were speciated using multiplex-PCR and evaluated for their acidogenic and aciduric potential.Results: Surprisingly, a high prevalence of Candida species (72.7%), either singly or in combination, was noted from both the proximal and occlusal cavities. C. tropicalis was the most prevalent species (47%; 34/72), followed by C. krusei (43.1%; 31/72) and C. albicans (40.3%; 29/72), with C. glabrata being the least (9.7%; 7/72). Over 45% low-pH niches (pH <7) of both sites yielded either dual or triple species of Candida. Genotyping revealed three distinct C. albicans genotypes (A, B, and C) with (14/29; 48.3%) of strains belonging to Genotype A. All four evaluated Candida species exhibited acidogenic and aciduric potential, C. tropicalis being the most potent.Conclusion: This, the first report of the high-density, multispecies, yeast colonization of deep-dentinal lesions in S-ECC, suggests that the Candida-biome plays a significant etiologic role in the condition, possibly due to their profound acidogenicity in milieus rich in dietary carbohydrates.

Clinical efficacy and the antimicrobial potential of silver formulations in arresting dental caries: a systematic review.

BACKGROUND: The use of silver-formulation as microbicide to arrest dentinal caries is gaining popularity. The primary objective of the present appraisal was to systematically review the clinical (in vivo) applications and antimicrobial potential of silver-containing formulations in arresting dentinal caries. Our secondary aim was to sum up the available in vitro applications of silver-containing formulations against cariogenic microbes isolated from dentine lesions. METHODS: Ovid MEDLINE, EBSCO host, Web of Science, and Cochrane Library databases was searched between January 2009-May 2019. RESULTS: In vivo: We observed conflicting evidence of antimicrobial efficacy of SDF on a diverse array of microbial taxa present in carious dentine of primary and permanent teeth. Moreover, there is insufficient evidence on the application of AgNP-fluoride as an effective microbicidal against cariogens of dentine lesions. In vitro: We found a good evidence of microbicidal efficacy of silver diamine fluoride (SDF) on selective cariogenic microbes in human dentine model. Additionally, a good evidence was noted of in vitro application of silver nanoparticles (AgNPs) as a useful microbicidal against S. mutans adhesion, growth and subsequent biofilm formation in human dentine models. CONCLUSIONS: Taken together, in vitro evidence indicates the promising antimicrobial potential of silver-based formulations (SDF and nanosilver) against the predominant cariogenic flora, particularly from dentine lesions. Post-treatment clinical data of either the bactericidal and bacteriostatic effects of SDF or nanosilver are sparse. Furthermore, the current understanding of the specific size, concentration, antimicrobial mechanisms, and toxicological aspects of nano-silver compounds is inadequate to draw firm conclusions on their clinical utility.

Silver diamine fluoride (SDF) used in childhood caries management has potent antifungal activity against oral Candida species.

BACKGROUND: The microbiome of Severe-Early Childhood Caries (S-ECC), is characterized by an ecosystem comprising bacterial and fungal species, with a predominance of Candida species. Hence, an anti-cariogen effective against both bacteria and fungi would be valuable in the management of S-ECC. Here we evaluate the antifungal effect of silver diamine fluoride (SDF) against 35-clinical yeast isolates (Ten-each of C. albicans, C. krusei, C. tropicalis and five C. glabrata strains) from dentinal caries-lesions from S-ECC. RESULTS: Disc-diffusion and time-kill assays as well as MIC50 and MIC90 evaluations against therapeutic concentrations confirmed the broad-spectrum anti-candidal potency of SDF. Ultrastructural images revealed morphologic aberrations of yeast-cell walls on exposure to SDF. All C. krusei and C. glabrata isolates were significantly more sensitive to SDF, relative to the standard antifungal fluconazole. Further, SDF appears to effectively abrogate filamentation of C. albicans even at very low concentrations. CONCLUSIONS: Our data, for the first time, elucidate the antifungal potency of SDF, in addition to its known antibacterial activity, in the management of S-ECC.

Candida biome of severe early childhood caries (S-ECC) and its cariogenic virulence traits.

The protected niche of deep-caries lesions is a distinctive ecosystem. We assessed the Candida biome and its cariogenic traits from dentin samples of 50 children with severe-early childhood caries (S-ECC). Asymptomatic, primary molars belonging to International Caries Detection and Assessment-ICDAS caries-code 5 and 6 were analyzed, and C. albicans (10-isolates), C. tropicalis (10), C. krusei (10), and C. glabrata (5) isolated from the lesions were then evaluated for their biofilm formation, acidogenicity, and the production of secreted hydrolases: hemolysins, phospholipase, proteinase and DNase. Candida were isolated from 14/43 ICDAS-5 lesions (32.5%) and 44/57 ICDAS-6 lesions (77.2%). Compared to, ICDAS-5, a significantly higher frequency of multi-species infestation was observed in ICDAS-6 lesions (p=0.001). All four candidal species (above) showed prolific biofilm growth, and an equal potency for tooth demineralization. A significant interspecies difference in the mean phospholipase, as well as proteinase activity was noted (p < 0.05), with C. albicans being the predominant hydrolase producer. Further, a positive correlation between phospholipase and proteinase activity of Candida-isolates was noted (r = 0.818, p < 0.001). Our data suggest that candidal mycobiota with their potent cariogenic traits may significantly contribute to the development and progression of S-ECC.

In vitro activity of Caspofungin combined with Fluconazole on mixed Candida albicans and Candida glabrata biofilm.

The objective of this study was to evaluate the antifungal effect of caspofungin (CAS) combined with fluconazole (FLU) on the biofilm biomass and cultivable viability and microstructure of Candida albicans and Candida glabrata mixed biofilm in vitro.Biofilms were formed in a 96-well microtiter plate for crystal violet assay and colony forming unit (CFU) method and grown on plastic coverslip disks for scanning electron microscopy. MIC50 of CAS and FLU against single Candida spp.and mixed Candida spp.biofilms were evaluated using crystal violet assay. Additional,C. albicans and C. glabrata mixed biofilms were incubated with subinhibitory CAS concentration plus FLU and their percentages of Candida biofilm reduction were calculated. We found that percentages of biofilm reduction were significantly decreased when CAS at 0.25MIC and FLU (0.25 or 0.5MIC) were combined (P< .05) but not different when CAS at 0.5 MIC combined with FLU at 0.25 or 0.5MIC, compared to CAS treatment alone. Structural analyses revealed that CAS/FLU combination-treated biofilms showed less hyphae and blastospores with some aberrant cells compared to control group. Although it was evident that a greater CFU of Candida glabrata were demonstrated in every group, the total viable cells derived from CAS/FLU combination-treated biofilms at any ratio were not significantly different from positive control. Overall, CAS/FLU combinations appeared to affect the quantity and cell architecture, but number of viable cell, of Candida albicans and Candida glabrata mixed biofilm. This antifungal effect was CAS concentration dependent.

Salivary cytokine profile in elders with Candida-related denture stomatitis.

OBJECTIVES: This study investigated risk factors of denture stomatitis, and the levels of cytokines in the saliva of elderly Candida-related denture stomatitis participants compared with adult individuals. MATERIAL AND METHODS: The occurrence of denture stomatitis in 128 patients with upper removable dentures was clinically examined. Participants were divided into two age groups as adult and elder. Risk factors associated with denture stomatitis were evaluated by questionnaire and oral and dental prosthesis examination. The quality of dentures was evaluated by direct examination. Palatal mucosa was swabbed for Candida carriage investigation, and whole unstimulated saliva was collected for cytokine detection. Salivary cytokines (interleukin (IL)-6, IL-8, IL-10, IL-17, intercellular adhesion molecule (ICAM)-1 and tumour necrosis factor (TNF)-α) were examined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The associations between the occurrence of denture stomatitis and either the quality of dentures or Candida isolation on palatal mucosa were significant. However, differences in the levels of salivary IL-6, IL-8, IL-10, IL-17, ICAM-1 and TNF-α between the denture wearers with and without denture stomatitis were undetectable. Adult and elderly Candida-related denture stomatitis patients also showed a similar level of salivary IL-6, IL-8, IL-10, IL-17, ICAM-1 and TNF-α. No correlation between the presence of denture stomatitis in the elder and the quantity of Candida infection was found. CONCLUSION: No association was found between the occurrence of Candida-related denture stomatitis and the concentrations of salivary IL-6, IL-8, IL-10, IL-17, ICAM-1 and TNF-α, regardless of age.