Mucormycosis of the Mandible and Tongue: A Systematic Scoping Review.

AIM: Mucormycosis is a rare human infection associated with Mucorales, a group of filamentous moulds found in different environmental niches. Its oral manifestations may occur in the mandible and tongue despite being rare. We aimed to systematically review the data on clinical manifestations, risk factors, diagnostic approaches, treatment options, and outcomes of mandibular and tongue mucormycosis. METHODS: An electronic search of articles published between January 1975 and November 2022 in PubMed, Web of Science, and EMBASE databases was performed. A total of 22 articles met the inclusion criteria and reported 27 cases of oral mucormycosis in total. RESULTS: Fourteen patients had mandibular mucormycosis signs unrelated to COVID-19 infection, 6 had SARS-CoV-2-related mandibular mucormycosis, and 6 had manifestations in the tongue. All published case reports during the COVID-19 pandemic were from India. Patient ages ranged from 4 months old to 82 years, and most patients had important comorbidities, such as blood dyscrasias related to immune deficiency and uncontrolled type 2 diabetes mellitus. The signs and symptoms of mandibular and tongue mucormycosis varied from dental pain, loose teeth, and nonhealing sockets to dysphagia and paraesthesia of the lip. Some patients also reported trismus, draining sinus tract, and facial pain. The diagnosis of oral mucormycosis was based on a combination of clinical, radiographic, and histopathologic findings by demonstrating fungal hyphae in tissue specimens. In most cases, mucormycosis was managed with systemic amphotericin B, strict glycaemic control, and aggressive surgical debridement of infected tissue, minimising the progression of the fungal infection and thus improving the survival rate. In some cases, combined antifungal therapy, antibiotic therapy, and chlorhexidine mouthwashes were used successfully. CONCLUSIONS: Recognition of the signs and symptoms by oral care providers is pertinent for the early diagnosis and treatment of tongue and mandibular mucormycosis, and providers should be aware of the possibility of this opportunistic fungal infection in patients with COVID-19. A multidisciplinary approach is recommended for the management of this lethal infection.

Probiotic Bifidobacteria in Managing Periodontal Disease: A Systematic Review.

Although various probiotic organisms have been evaluated for their utility in the management of periodontitis, their strain-specific mechanisms of action are still unclear. We aimed to systematically review the effect of bifidobacterial probiotics on periodontopathogens and host immune responses in periodontal diseases. An electronic search of articles published until June 2022 in Medline, PubMed, Web of Science, and Cochrane Library databases was performed. Randomised controlled trials (RCTs) and in vitro and animal studies were assessed, and the data regarding antimicrobial properties, immunomodulation, and clinical outcomes were analysed. A total of 304 studies were screened, but only 3 RCTs and 6 animal and in vitro studies met the inclusion criteria. The use of different strains of bifidobacteria led to (1) a reduction of key players of the red complex periodontopathogens; (2) reduced levels of pro-inflammatory cytokines (eg, interleukin [IL]1-ß and IL-8) and higher levels of anti-inflammatory cytokines (IL-10); (3) enhanced levels of osteoprotegerin and reduced levels of receptor activator of nuclear factor kappa-B ligand; and (4) a reduction of the dental plaque, bleeding on probing, alveolar bone loss, and clinical attachment loss. Bifidobacterial probiotic adjuvant supplementation, especially with Bifidobacterium animalis subspecies lactis, appears to help improve clinical periodontal parameters and develop a healthy plaque microbiome through microbiological and immunomodulatory pathways. Further human and animal studies are warranted prior to the therapeutic use of bifidobacteria in the routine management of periodontal infections.

Orofacial Mycoses in Coronavirus Disease-2019 (COVID-19): A Systematic Review.

OBJECTIVES: Studies reviewing orofacial mycoses in coronavirus disease-2019 (COVID-19) caused by severe acute respiratory syndrome 2 (SARS-CoV-2) infection are sparse. Here we review the major oral and maxillofacial mycoses of COVID-19, the associated comorbidities, and the probable precipitating factors. METHODS: English-language manuscripts published between March 2020 and October 2021 were searched using PubMed, OVID, SCOPUS, and Web of Science databases, using appropriate keywords. RESULTS: We identified 30 articles across 14 countries, which met the inclusion criteria of PRISMA guidelines. These yielded a total of 292 patients with laboratory-confirmed COVID-19, 51.4% (n = 150) of whom presented with oral and maxillofacial fungal infections, mainly comprising candidosis, mucormycosis, and aspergillosis. Candida infections were the most prevalent, present in 64% (n = 96), followed by mucormycosis, and only a single case of aspergillosis was noted. Oral and maxillofacial mycoses were predominantly seen in those with comorbidities, especially in those with diabetes (52.4%). Oral mucormycosis was noted in 8.6% (n = 13) and mainly manifested on the hard palate. An overall event rate of oral/maxillofacial mucormycosis manifestation in patients with COVID-19 with diabetes mellitus type 1/2 was about 94% (49/52; 95% confidence interval, 0.73%-0.89%), implying a very high association between diabetes mellitus and the latter condition. All fungal infections appeared either concurrently with COVID-19 symptoms or during the immediate recovery period. CONCLUSIONS: SARS-CoV-2 infection-related immunosuppression, steroid therapy, as well as comorbidities such as diabetic hyperglycemia appear to be the major predisposing factors for the onset of oral and maxillofacial mycoses in patients with COVID-19 across all age groups.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) viral positivity and their burden in saliva of asymptomatic carriers - a systematic review and meta-analysis.

AIMS: The coronavirus disease 2019 (COVID-19) due to the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) can present either as an asymptomatic carrier state or an acute respiratory disease, with or without severe pneumonia. The asymptomatic carriers are a challenge for the dental profession as the infection could be transmitted via virus-laden, and saliva in dental settings through aerosol-generating procedures (AGPSs). The aim of this review was to perform a systematic review of SARS-CoV-2 in the saliva of asymptomatic individuals. MATERIALS AND METHODS: PubMed, Google scholar, and MedRxiv databases were searched between and a systematic review and meta-analysis of the available data were performed to assess the viral burden in the saliva of asymptomatic carriers of SARS-CoV-2. All investigators of the included studies used qRT-PCR to detect SARS-CoV-2 and yield quantitative data (the Ct values) appertaining to the viral load. RESULTS: A total of 322 records in the English literature were identified, and eight studies with 2642 SARS-CoV-2-positive and asymptomatic individuals were included in the final analysis. Of these, 16.7% (95% CI: 11-23%) yielded SARS-CoV-2-positive saliva samples in comparison to 13.1% (95% CI: 12-17%) of the respiratory specimens (nasopharyngeal or nose-throat swabs). CONCLUSION: As approximately 1 in 5 to 1 in 10 asymptomatic individuals harbour SARS-CoV-2 in either saliva or respiratory secretions, our results highlight the need for continued vigilance and the critical importance of maintaining strict, additional infection control regimens for the foreseeable future to mitigate the potential risks of COVID-19 transmission in dentistry.

Tea extracts inhibit the attachment of streptococci to oral/dental substrata by reducing hydrogen bonding energies.

Previous work in the authors' lab demonstrated that tea extracts significantly suppressed streptococcal colonization of abiotic substrata by coating the bacterial cell surfaces with tea components. In this study, the physico-chemical mechanisms by which the tea coating inhibits cellular attachment are demonstrated. The changes in the cell surface physico-chemical properties of streptococci, induced by tea extracts, were measured. Using these results, surface interaction energies were calculated between streptococcal cells and hard surfaces (glass, stainless steel, hydroxyapatite and titanium) within the cellular attachment system exploiting the extended Derjaguin-Landau-Verwey-Overbeek theory. The net energy outcomes were compared with experiment results of attachment assays to validate the predictability of the model. The results showed that the tea extracts inhibited the attachment of the bacteria by 11.1%-91.5%, and reduced the interaction energy by 15.4%-94.9%. It was also demonstrated that the abilities of the bacteria to attach to hard surfaces correlated well with their net interaction energies. The predominant interaction in the systems was found to be hydrogen bonding. In conclusion, tea extracts suppress streptococcal attachment to hard substrata by limiting the formation of hydrogen bonds.

Comparative Metabolomics Reveals the Microenvironment of Common T-Helper Cells and Differential Immune Cells Linked to Unique Periapical Lesions.

Periapical abscesses, radicular cysts, and periapical granulomas are the most frequently identified pathological lesions in the alveolar bone. While little is known about the initiation and progression of these conditions, the metabolic environment and the related immunological behaviors were examined for the first time to model the development of each pathological condition. Metabolites were extracted from each lesion and profiled using gas chromatography-mass spectrometry in comparison with healthy pulp tissue. The metabolites were clustered and linked to their related immune cell fractions. Clusters I and J in the periapical abscess upregulated the expression of MMP-9, IL-8, CYP4F3, and VEGF, while clusters L and M were related to lipophagy and apoptosis in radicular cyst, and cluster P in periapical granuloma, which contains L-(+)-lactic acid and ethylene glycol, was related to granuloma formation. Oleic acid, 17-octadecynoic acid, 1-nonadecene, and L-(+)-lactic acid were significantly the highest unique metabolites in healthy pulp tissue, periapical abscess, radicular cyst, and periapical granuloma, respectively. The correlated enriched metabolic pathways were identified, and the related active genes were predicted. Glutamatergic synapse (16-20),-hydroxyeicosatetraenoic acids, lipophagy, and retinoid X receptor coupled with vitamin D receptor were the most significantly enriched pathways in healthy control, abscess, cyst, and granuloma, respectively. Compared with the healthy control, significant upregulation in the gene expression of CYP4F3, VEGF, IL-8, TLR2 (P < 0.0001), and MMP-9 (P < 0.001) was found in the abscesses. While IL-12A was significantly upregulated in cysts (P < 0.01), IL-17A represents the highest significantly upregulated gene in granulomas (P < 0.0001). From the predicted active genes, CIBERSORT suggested the presence of natural killer cells, dendritic cells, pro-inflammatory M1 macrophages, and anti-inflammatory M2 macrophages in different proportions. In addition, the single nucleotide polymorphisms related to IL-10, IL-12A, and IL-17D genes were shown to be associated with periapical lesions and other oral lesions. Collectively, the unique metabolism and related immune response shape up an environment that initiates and maintains the existence and progression of these oral lesions, suggesting an important role in diagnosis and effective targeted therapy.

Prevalence of unculturable bacteria in the periapical abscess: A systematic review and meta-analysis.

OBJECTIVE: To assess the prevalence of unculturable bacteria in periapical abscess, radicular cyst, and periapical granuloma. METHODS: PubMed, Scopus, Science Direct, and Ovid databases were systematically searched from January 1990 to May 2020. All the included studies were cross-sectional design. The risk of bias was assessed using Joanna Briggs Institute check-list. Heterogeneity was described using meta-regression and mixed-effects model for lesion, country, and sequence technique moderators. Funnel plot and unweighted Egger's regression test were used to estimate the publication bias. Microbiome data on diversity, abundance, and frequency of unculturable bacteria in the periapical lesions were reviewed, analysed, and the principal component analysis (PCA) was performed. RESULTS: A total of 13 studies out of 14,780, were selected for the final analysis. These studies focused on the prevalence of unculturable bacteria in periapical abscesses and related lesions. Approximately 13% (95% CI: 7-23%) of the cumulative number of bacteria derived from periapical abscesses was unculturable. Country moderator significantly (P = 0.05) affects the diversity summary proportion. While the pooled frequency of unculturable bacteria was 8%; 95% CI: 5, 14%, the estimate of the pooled abundance of unculturable bacteria was 5%; 95% CI: 2, 12% with a significant (P = 0.05) country moderator that affects the abundance summary proportion. Of the 62 unculturable bacteria, 35 were subjected to PCA and Peptostreptococcus sp. oral clone CK035 was the most abundant species in periapical abscesses. Hybridization techniques were found to be the most reliable molecular methods in detecting the abundance and frequency of unculturable bacteria. CONCLUSION: The significant prevalence of unculturable bacteria in the periapical abscess, suggests that they are likely to play, a yet unknown, critical role in the pathogenesis and progression of the disease. Further research remains to be done to confirm their specific contributions in the virulence and disease progression.

Tea extracts modulate oral biofilm development by altering bacterial hydrophobicity and aggregation.

OBJECTIVES: This study aims to investigate the effects of tea extracts on biofilm formation by oral streptococci and the potential mechanisms behind the effects. DESIGN: We examined the effects of five types of tea extracts (green, oolong, black, pu-erh and chrysanthemum tea) on cell surface hydrophobicity and auto-aggregation of three different streptococcal species (Streptococcus mutans, Streptococcus salivarius and Streptococcus mitis) and evaluated their biofilm formation on four disparate hard surfaces (glass, stainless steel, hydroxyapatite and titanium). The correlation between biofilm formation and the cellular properties were investigated in order to study the mechanisms by which the tea extracts affect biofilm formation. RESULTS: Results show that the tea extracts reduced cell surface hydrophobicity (by up to 57.9 %) and, in some cases, altered cellular auto-aggregation (by up to 12 %) and biofilm formation (by up to 2.61 log CFU cm-2). Specifically, oolong tea extract was found to enhance biofilm formation by increasing cellular auto-aggregation and pu-erh tea extract retarded biofilm formation by increasing auto-aggregation. Biofilm formation correlated well to cell surface hydrophobicity and auto-aggregation in combination, but not to either one alone as determined by multiple linear regression analysis. CONCLUSIONS: Tea extracts have the ability to modulate streptococcal biofilm formation by altering cell surface hydrophobicity and cellular aggregation.

Plant components affect bacterial biofilms development by altering their cell surface physicochemical properties: a predictability study using Actinomyces naeslundii.

We hypothesized that the initial events leading to biofilm formation by bacteria, in general, are predominantly mediated by cell surface physicochemical interactions, and that natural products can impact the process by altering cell surface physicochemical properties. We exemplified this phenomenon using Actinomyces naeslundii as the model organism, and using tea products to modify its cell surface physicochemical properties. To test the hypothesis, a non-linear multiple regression model incorporating a normal distribution curve was constructed to explain the impact of tea extracts on the physiochemical processes of biofilm formation by A. naeslundii. The model utilized tea extract-induced changes in cell surface physicochemical properties as independent variables, and the corresponding biofilm formation as a dependent variable. Five different tea extracts were used to treat A. naeslundii, and their impact on the cell surface hydrophobicity, charge, auto-aggregation, attachment and biofilm formation on four different hard surfaces were measured and the data were used to construct the model. The established model was then tested in independent experiments involving other plant extracts and purified phytochemicals. Experimental results showed that the tea extracts significantly reduced cell surface hydrophobicity (by up to 21.3%), increased cell surface charge and auto-aggregation (by up to 4.5 mV and 14.9%, respectively), inhibited attachment (by 0.6-2.5 log CFU cm-2) and affected biofilm formation (by up to 0.6 log CFU cm-2). The model indicated that both cell surface hydrophobicity and charge played an important role in bacterial auto-aggregation and attachment, and that the latter two phenomena significantly correlated with subsequent biofilm development. The accuracy of the model construct was approximately 64%. This modelling approach can be employed for other microbial colonization systems to predict biofilm formation, and to study the impact of cell surface physicochemical properties in biofilm development.

Viral, bacterial, and fungal infections of the oral mucosa: Types, incidence, predisposing factors, diagnostic algorithms, and management.

For millions of years, microbiota residing within us, including those in the oral cavity, coexisted in a harmonious symbiotic fashion that provided a quintessential foundation for human health. It is now clear that disruption of such a healthy relationship leading to microbial dysbiosis causes a wide array of infections, ranging from localized, mild, superficial infections to deep, disseminated life-threatening diseases. With recent advances in research, diagnostics, and improved surveillance we are witnessing an array of emerging and re-emerging oral infections and orofacial manifestations of systemic infections. Orofacial infections may cause significant discomfort to the patients and unnecessary economic burden. Thus, the early recognition of such infections is paramount for holistic patient management, and oral clinicians have a critical role in recognizing, diagnosing, managing, and preventing either new or old orofacial infections. This paper aims to provide an update on current understanding of well-established and emerging viral, bacterial, and fungal infections manifesting in the human oral cavity.

Oral Candida carriage among cigarette- and waterpipe-smokers, and electronic cigarette users.

OBJECTIVE: The aim of this cross-sectional study was to compare oral Candida carriage among cigarette- and waterpipe-smokers, electronic cigarette (E-Cig) users, and never-smokers. METHODS: Demographic data and information regarding smoking and vaping were collected using a questionnaire. Number of missing teeth and unstimulated whole salivary flow rate (UWSFR) were recorded. Oral Candida samples were collected and identified using concentrated oral rinse culture technique and PCR, respectively. Level of significance was set at p < 0.05. RESULTS: Thirty-four cigarette-smokers (Group-1), 33 waterpipe-smokers (Group-2), 30 E-Cig users (Group-3), and 32 never-smokers (Group-4) were included. All were male participants with comparable mean ages. In groups 1, 2, 3, and 4, oral Candida carriage rate was 100%, 100%, 83.3% and 50%, respectively. The most commonly isolated oral yeast species in all groups was C. albicans. C. albicans carriage was significantly higher in groups 1 (p < 0.05), 2 (p < 0.05), and 3 (p < 0.05) than Group-4. Data stratification for age, missing teeth, and UWSFR showed no significant difference in oral yeasts carriage in groups 1, 2, and 3. CONCLUSIONS: Oral C. albicans carriage was significantly higher among cigarette- and waterpipe-smokers and E-Cig users than never-smokers. No significant differences were identified among groups in the oral carriage of other Candida species.

The Effect of Nutritive and Non-Nutritive Sweeteners on the Growth, Adhesion, and Biofilm Formation of Candida albicans and Candida tropicalis.

OBJECTIVE: To determine the effect of glucose, sucrose, and saccharin on growth, adhesion, and biofilm formation of Candida albicans and Candida tropicalis. MATERIALS AND METHODS: The growth rates of mono-cultures of planktonic C. albicans and C. tropicalis and 1:1 mixed co-cultures were determined in yeast nitrogen broth supplemented with 5% (30 mM) and 10% (60 mM) glucose, sucrose, and saccharin, using optical density measurements at 2-h intervals over a 14-h period. Adhesion and biofilm growth were performed and the growth quantified using a standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The biofilm architecture was visualized using scanning electron microscopy. One- and two-way analysis of variance (ANOVA) was performed to analyse the differences among multiple means. RESULTS: The highest planktonic growth was noted in 5% glucose after 14 h (p < 0.05). No significant planktonic growth was observed in either concentration of saccharin. Both the concentrations of glucose and sucrose elicited significantly increased adhesion from MTT activity of 0.017 to >0.019 in mono- as well as co-cultures (p < 0.05), whilst the lower concentration of saccharin significantly dampened the adhesion. Maximal biofilm growth was observed in both species with the lower concentration of sucrose (5%), although a similar concentration of saccharin abrogated biofilm development: the highest MTT value (>0.35) was obtained for glucose and the lowest (>0.15) for saccharin. CONCLUSION: In this study, glucose and sucrose accelerated the growth, adhesion, and biofilm formation of Candida species. However, the non-nutritive sweetener saccharin appeared to dampen, and in some instances suppress, these virulent attributes of Candida.

Effects of tea extracts on the colonization behaviour of Candida species: attachment inhibition and biofilm enhancement.

Purpose. We assessed the effects of four different types of tea extracts (green, oolong, black and pu-erh tea) on cellular surface properties (hydrophobicity and auto-aggregation) and the colonization attributes (attachment and biofilm formation) of four strains of Candida albicans and three strains of Candida krusei.Methodology. The cellular surface properties were determined using spectrophotometry. The colonization activities were quantified using colorimetric viability assays and visualized using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM).Results. The tea extracts, in general, reduced the hydrophobicity (by 8-66 %) and auto-aggregation (by 20-65 %), and inhibited the attachment of two C. krusei strains (by 41-88 %). Tea extracts enhanced the biofilm formation of one C. albicans and two C. krusei strains (by 1.4-7.5-fold). The observed reduction in hydrophobicity strongly correlated with the reduction in attachment of the two C. krusei strains (P<0.05). The ultrastructural images of the tea-treated C. krusei biofilm cells demonstrated central indentations, although they remained viable.Conclusion. The tea extracts have the ability to retard C. krusei adhesion to glass surfaces, possibly by reducing fungal cellular hydrophobicity, whilst paradoxically promoting biofilm formation. In practical terms, therefore, consumption of tea beverages appears to have a complex effect on oral candidal colonization.

Culture media profoundly affect Candida albicans and Candida tropicalis growth, adhesion and biofilm development

As there are sparse data on the impact of growth media on the phenomenon of biofilm development for Candida we evaluated the efficacy of three culture media on growth, adhesion and biofilm formation of two pathogenic yeasts, Candida albicans and Candida tropicalis. The planktonic phase yeast growth, either as monocultures or mixed cultures, in sabouraud dextrose broth (SDB), yeast nitrogen base (YNB), and RPMI 1640 was compared, and adhesion as well as biofilm formation were monitored using MTT and crystal violet (CV) assays and scanning electron microscopy. Planktonic cells of C. albicans, C. tropicalis and their 1:1 co-culture showed maximal growth in SDB. C. albicans/C. tropicalis adhesion was significantly facilitated in RPMI 1640 although the YNB elicited the maximum growth for C. tropicalis. Similarly, the biofilm growth was uniformly higher for both species in RPMI 1640, and C. tropicalis was the slower biofilm former in all three media. Scanning electron microscopy images tended to confirm the results of MTT and CV assay. Taken together, our data indicate that researchers should pay heed to the choice of laboratory culture media when comparing relative planktonic/biofilm growth of Candida. There is also a need for standardisation of biofilm development media so as to facilitate cross comparisons between laboratories.

Culture media profoundly affect Candida albicans and Candida tropicalis growth, adhesion and biofilm development.

As there are sparse data on the impact of growth media on the phenomenon of biofilm development for Candida we evaluated the efficacy of three culture media on growth, adhesion and biofilm formation of two pathogenic yeasts, Candida albicans and Candida tropicalis. The planktonic phase yeast growth, either as monocultures or mixed cultures, in sabouraud dextrose broth (SDB), yeast nitrogen base (YNB), and RPMI 1640 was compared, and adhesion as well as biofilm formation were monitored using MTT and crystal violet (CV) assays and scanning electron microscopy. Planktonic cells of C. albicans, C. tropicalis and their 1:1 co-culture showed maximal growth in SDB. C. albicans/C. tropicalis adhesion was significantly facilitated in RPMI 1640 although the YNB elicited the maximum growth for C. tropicalis. Similarly, the biofilm growth was uniformly higher for both species in RPMI 1640, and C. tropicalis was the slower biofilm former in all three media. Scanning electron microscopy images tended to confirm the results of MTT and CV assay. Taken together, our data indicate that researchers should pay heed to the choice of laboratory culture media when comparing relative planktonic/biofilm growth of Candida. There is also a need for standardisation of biofilm development media so as to facilitate cross comparisons between laboratories.

Enterococcus Species in the Oral Cavity: Prevalence, Virulence Factors and Antimicrobial Susceptibility.

Enterococci are considered as transient constituent components of the oral microbiome that may cause a variety of oral and systemic infections. As there is sparse data on the oral enterococcal prevalence, we evaluated the Enterococcus spp. and their virulence attributes including antimicrobial resistance in a healthy Brazilian cohort. A total of 240 individuals in different age groups were studied (children 4-11 yrs, adolescents 12-17 yrs, young adults 18-29 yrs, adults 30-59 yrs, elderly over 60 yrs). Oral rinses were collected and isolates were identified by API 20 Strep and confirmed by 16S rDNA sequencing. E. faecalis isolates, in particular, were evaluated for virulence attributes such as their biofilm formation potential, and susceptibility to antimicrobials and an antiseptic, chlorhexidine gluconate. A total of 40 individuals (16.6%) and 10% children, 4% adolescents, 14% young adults, 30% adults, and 25% elderly carried oral enterococci. The oral enterococcal burden in adolescents was significantly lower than in the adults (p = 0.000) and elderly (p = 0.004). The proportion of carriers was higher among females (p = 0.001). E. faecalis was the most frequent isolate in all the age groups (p = 0.000), followed by E. durans and E. faecium. Whilst all the clinical isolates were able to form biofilms, only a proportion of them were able to produce lipase (92%), hemolysin (38%), and gelatinase (39%). Of all the isolates 53.8% were resistant to tetracycline, 12.3% to amoxicillin, 16.0% to ampicillin, 20.8% to chloramphenicol and 43.4% to erythromycin. None of the isolates were resistant to vancomycin. Our data suggest that in this Brazilian cohort the oral cavity may act as a significant reservoir of rather virulent and antibiotic resistant enterococci, with an increasing degree of carriage in the adults and elderly. Hence clinicians should be cognizant of this silent reservoir of virulent enterococci that may pose a particular threat of nosocomial infection.

Probiotic lactobacilli inhibit early stages of Candida albicans biofilm development by reducing their growth, cell adhesion, and filamentation.

We evaluated the inhibitory effects of the probiotic Lactobacillus species on different phases of Candida albicans biofilm development. Quantification of biofilm growth and ultrastructural analyses were performed on C. albicans biofilms treated with Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus acidophilus planktonic cell suspensions as well as their supernatants. Planktonic lactobacilli induced a significant reduction (p < 0.05) in the number of biofilm cells (25.5-61.8 %) depending on the probiotic strain and the biofilm phase. L. rhamnosus supernatants had no significant effect on the mature biofilm (p > 0.05), but significantly reduced the early stages of Candida biofilm formation (p < 0.01). Microscopic analyses revealed that L. rhamnosus suspensions reduced Candida hyphal differentiation, leading to a predominance of budding growth. All lactobacilli negatively impacted C. albicans yeast-to-hyphae differentiation and biofilm formation. The inhibitory effects of the probiotic Lactobacillus on C. albicans entailed both cell-cell interactions and secretion of exometabolites that may impact on pathogenic attributes associated with C. albicans colonization on host surfaces and yeast filamentation. This study clarifies, for the first time, the mechanics of how Lactobacillus species may antagonize C. albicans host colonization. Our data elucidate the inhibitory mechanisms that define the probiotic candicidal activity of lactobacilli, thus supporting their utility as an adjunctive therapeutic mode against mucosal candidal infections.

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. Candida 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, Candida 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.

Probiotics as Antifungals in Mucosal Candidiasis.

Candidais an opportunistic pathogen that causes mucosal and deep systemic candidiasis. The emergence of drug resistance and the side effects of currently available antifungals have restricted their use as long-term prophylactic agents for candidal infections. Given this scenario, probiotics have been suggested as a useful alternative for the management of candidiasis. We analyzed the available data on the efficacy of probiotics in candidal colonization of host surfaces. A number of well-controlled studies indicate that probiotics, particularly lactobacilli, suppressCandidagrowth and biofilm development in vitro.A few clinical trials have also shown the beneficial effects of probiotics in reducing oral, vaginal, and enteric colonization byCandida; alleviation of clinical signs and symptoms; and, in some cases, reducing the incidence of invasive fungal infection in critically ill patients. Probiotics may serve in the future as a worthy ally in the battle against chronic mucosal candidal infections.

The yield and quality of cellular and bacterial DNA extracts from human oral rinse samples are variably affected by the cell lysis methodology.

Recent culture-independent studies have enabled detailed mapping of human microbiome that has not been hitherto achievable by culture-based methods. DNA extraction is a key element of bacterial culture-independent studies that critically impacts on the outcome of the detected microbial profile. Despite the variations in DNA extraction methods described in the literature, no standardized technique is available for the purpose of microbiome profiling. Hence, standardization of DNA extraction methods is urgently needed to yield comparable data from different studies. We examined the effect of eight different cell lysis protocols on the yield and quality of the extracted DNA from oral rinse samples. These samples were exposed to cell lysis techniques based on enzymatic, mechanical, and a combination of enzymatic-mechanical methods. The outcome measures evaluated were total bacterial population, Firmicutes levels and human DNA contamination (in terms of surrogate GAPDH levels). We noted that all three parameters were significantly affected by the method of cell lysis employed. Although the highest yield of gDNA was obtained using lysozyme-achromopeptidase method, the lysozyme-zirconium beads method yielded the peak quantity of total bacterial DNA and Firmicutes with a lower degree of GAPDH contamination compared with the other methods. Taken together our data clearly points to an urgent need for a consensus, standardized DNA extraction technique to evaluate the oral microbiome using oral rinse samples. Further, if Firmicutes levels are the focus of investigation in oral rinse microbiome analyses then the lysozyme-zirconium bead method would be the method of choice in preference to others.