Diversity of site-specific microbes of occlusal and proximal lesions in severe- early childhood caries (S-ECC)

Background: Severe-early childhood caries (S-ECC) a global problem of significant concern, commonly manifest on the occlusal, and proximal surfaces of affected teeth. Despite the major ecological differences between these two niches the compositional differences, if any, in the microbiota of such lesions is unknown. Methods: Deep-dentine caries samples from asymptomatic primary molars of children with S-ECC (n 19) belonging to caries-code 5/6, (ICDAS classification) were evaluated. Employing two primer pools, we amplified and compared the bacterial 16S rRNA gene sequences of the seven hypervariable regions (V2—V4 and V6—V9) using NGS-based assay. Results: Bray-Curtisevaluation indicated that occlusal lesions (OL) had a more homogeneous community than the proximal lesions (PL) with significant compositional differences at the species level (p = 0.01; R- 0.513). Together, the occlusal and proximal niches harbored 263 species, of which 202 (76.8%) species were common to both, while 49 (18.6%) and 12 (4.6%) disparate species were exclusively isolated from the proximal and occlusal niches, respectively. The most commonl genera at both niches included Streptococcus, Prevotella, and Lactobacillus. S. mutans was predominant in PL (p ≤ 0.05), and Atopobium parvulum (p = 0.01) was predominant in OL. Conclusions: Distinct differences exist between the caries microbiota of occlusal and proximal caries in S-ECC

Biodiversity of the human oral mycobiome in health and disease

The organisms that colonize the human body over a lifetime are diverse, extensive and gargantuan. A fair proportion of the microbiota that constitutes this human microbiome live within our oral cavities mostly as harmonious associates causing only sporadic disease. An important core constituent of the microbiome is the mycobiome, representing various fungal genera. Up until recently, only a few species of fungi, mainly Candida species, were thought to constitute the human oral mycobiome. The reasons for this are manifold, although the uncultivable nature of many fungi in conventional laboratory media, and their complex genetic composition seem to be the major factors which eluded their detection over the years. Nevertheless, recent advances in computing and high‐throughput sequencing such as next‐generation sequencing (NGS) platforms have provided us a panoramic view of a totally new world of fungi that are human oral cohabitués. Their diversity is perplexing, and functionality yet to be deciphered. Here, we provide a glimpse of what is currently known of the oral mycobiome, in health and disease, with some future perspectives

Candida albicans orf19.3727 encodes phytase activity and is essential for human tissue damage.

Candida albicans is a clinically important human fungal pathogen. We previously identified the presence of cell-associated phytase activity in C. albicans. Here, we reveal for the first time, that orf19.3727 contributes to phytase activity in C. albicans and ultimately to its virulence potency. Compared with its wild type counterpart, disruption of C. albicans orf19.3727 led to decreased phytase activity, reduced ability to form hyphae, attenuated in vitro adhesion, and reduced ability to penetrate human epithelium, which are the major virulence attributes of this yeast. Thus, orf19.3727 of C. albicans plays a key role in fungal pathogenesis. Further, our data uncover a putative novel strategy for anti-Candidal drug design through inhibition of phytase activity of this common pathogen

The effect of the combined use of silver diamine fluoride and potassium iodide in disrupting the plaque biofilm microbiome and alleviating tooth discoloration: A systematic review.

Silver diamine fluoride (SDF) is used in minimally invasive dentistry for arresting dental caries. However, discoloration of teeth is a significant side effect that has limited the use of SDF. Hence, the application of potassium iodide (KI) following SDF has been proposed to ameliorate the staining. Although antimicrobial activity is one of the major mechanisms of the caries-arresting effect of SDF, the antimicrobial potency of SDF/KI combination is unclear. Thus, the primary objective of this systematic review was to appraise the studies on the antimicrobial efficacy of SDF/KI combination on cariogenic microbes. The secondary objective was to summarize the evidence on the potential of KI in reducing the discoloration associated with the application of SDF. Electronic databases of Medline via PubMed, Cochrane Library, Web of Science, and EBSCO host were searched for English language manuscripts from January 2005 to 15th November 2020. The reference lists of these manuscripts were manually searched for additional studies. Twelve studies were included in the final analysis, seven of which have investigated the antimicrobial efficacy of SDF/KI, and the rest have examined the anti-staining potential of KI. The exploratory findings from the reviewed articles revealed the promising antimicrobial potential of SDF/KI on cariogenic microbes associated with dentine caries. There is, however, contradictory evidence on the effect of SDF/KI on tooth color. The reviewed in-vitro studies indicated significant effectiveness of KI in preventing staining. A clinical trial on primary dentition showed 25% reduction in the incidence of staining by SDF after applying KI, while a clinical study on root caries in adults showed no significant effect. Within the methodological limitations of this review, we conclude that for arresting dental caries, SDF could be combined with KI, as there may be a lower likelihood of staining. Further, well-designed clinical trials on the antimicrobial and anti-staining effect of SDF/KI are needed to obtain more robust evidence

Caspofungin-induced in-vitro post-antifungal effect and its impact on adhesion related traits of oral Candida dubliniensis and Candida albicans isolates

Adhesion to buccal epithelial cells (BEC) and denture acrylic surfaces (DAS), germ tube (GT) formation and cell surface hydrophobicity (CSH) are all virulence traits involved in the pathogenicity of Candida. Post-antifungal effect (PAFE) also have a bearing on pathogenicity and virulence of Candida. Candida dubliniensis is associated with oral and systemic candidosis, which can be managed with caspofungin. There is no published information on caspofungin-induced PAFE and its impact on adhesion traits of C. dubliniensis isolates. Thus, the purpose of this investigation was to determine the in vitro duration of PAFE on 20 C. dubliniensis isolates following transient exposure to caspofungin. Furthermore the impacts of caspofungin-induced PAFE on adhesion to BEC and DAS, GT formation and CSH of these isolates were also determined. After establishing the minimum inhibitory concentration (MIC) of caspofungin, C. dubliniensis isolates were exposed to sub-lethal concentrations (×3MIC) of caspofungin for 1hr. Thereafter the duration of PAFE, adhesion to BEC and DAS, GT formation and CSH were determined by previously described in-vitro assays. MIC (μg/mL) of C. dubliniensis isolates to caspofungin ranged from 0.004 to 0.19. Caspofungin-induced mean PAFE on C. dubliniensis isolates was 2.17hr. Exposure to caspofungin suppressed the ability of C. dubliniensis isolates to adhere to BEC and DAS, form GT and CSH by 69.97%, 71.95%, 90.06% and 32.29% (

Antifungal susceptibility in serum and virulence determinants of Candida bloodstream isolates from Hong Kong

Candida bloodstream infections (CBI) are one of the most common nosocomial infections globally, and they account for a high mortality rate. The increasing global prevalence of drug-resistant Candida strains has also been posing a challenge to clinicians. In this study, we comprehensively evaluated the biofilm formation and production of hemolysin and proteinase of 63 CBI isolates derived from a hospital setting in Hong Kong as well as their antifungal susceptibility both in the presence and in the absence of human serum, using standard methodology. C. albicans was the predominant species among the 63 CBI isolates collected, and non-albicans Candida species accounted for approximately one third of the isolates (36.5%). Of them, C. tropicalis was the most common non-albicans Candida species. A high proportion (31.7%) of the CBI isolates (40% of C. albicans isolates, 10% of C. tropicalis isolates, 11% of C. parapsilosis isolates and 100% of C. glabrata isolates) were found to be resistant to fluconazole. One of the isolates (C. tropicalis) was resistant to amphotericin B. A rising prevalence of drug-resistance CBI isolates in Hong Kong was observed with reference to a previous study. Notably, all non-albicans Candida species, showed increased hemolytic activity relative to C. albicans, whilst C. albicans, C. tropicalis and C. parapsilosis exhibited proteinase activities. Majority of the isolates were capable of forming mature biofilms. Interestingly, the presence of serum distorted the yeast sensitivity to fluconazole, but not amphotericin B. Taken together, our findings demonstrate that CBI isolates of Candida have the potential to express to varying extent their virulence attributes (e.g. biofilm formation, hemolysin production and proteinase activity) and these, together with perturbations in their antifungal sensitivity in the presence of serum, may contribute to treatment complication in candidemia. The effect of serum on antifungal activity warrants further investigations, as it has direct clinical relevance to the treatment outcome in subjects with candidemia

Fluoride and silver concentrations of silver diammine fluoride solutions for dental use

Objective: To determine the short-term stability of free fluoride ion concentrations and acidity (pH values) of three commercially available SDF solutions over time. Methods: Three SDF products for caries control were studied: Cariestop-12%, Cariestop-30% and Saforide-38%. Their expected fluoride ion concentrations were 14,200, 35,400 and 44,800 ppm, respectively. The fluoride ion concentrations were determined with an ion-selective electrode. The acidity was determined with a pH electrode. The measurements were performed when open and at 7 and 28 days. Results: The mean fluoride ion concentrations of the freshly opened bottles were 12,525 ± 450, 13,200 ± 2060 and 55,800 ± 2536 ppm, respectively. The mean pH values were 9.4 ± 0.1, 10.4 ± 0.1 and 10.2 ± 0.2, respectively. No significant change (P > 0.05) in the fluoride ion concentrations or the acidity was detected after 7 or 28 days. Conclusion: The three SDF tested solutions were alkaline. The fluoride ion concentrations of Cariestop-30% and Saforide-38% were considerably different. The fluoride ion concentrations and acidity of the products demonstrated a short-term stability over 28 days

Determination of lactate dehydrogenase (LDH) from the culture supernatants of the RHE tissues upon fungal invasion.

<p>The amount of released LDH was determined using CytoTox 96 nonradioactive cytotoxicity assay (n = 3; *<i>p</i> < 0.05).</p