scholarly journals Intervening in Symbiotic Cross-Kingdom Biofilm Interactions: a Binding Mechanism-Based Nonmicrobicidal Approach

mBio ◽  
2021 ◽  
Vol 12 (3) ◽  
Author(s):  
H. E. Kim ◽  
A. Dhall ◽  
Y. Liu ◽  
M. Bawazir ◽  
H. Koo ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Streptococcus Mutans ◽  
Single Molecule ◽  
Mechanical Stability ◽  
Tooth Enamel ◽  
Oral Disease ◽  
Enzyme Treatment ◽  
Human Tooth ◽  
Tooth Decay ◽  
Content Type

ABSTRACT Early childhood caries is a severe oral disease that results in aggressive tooth decay. Particularly, a synergistic association between a fungus, Candida albicans, and a cariogenic bacterium, Streptococcus mutans, promotes the development of hard-to-remove and highly acidic biofilms, exacerbating the virulent damage. These interactions are largely mediated via glucosyltransferases (GtfB) binding to mannans on the cell wall of C. albicans. Here, we present an enzymatic approach to target GtfB-mannan interactions in this cross-kingdom consortium using mannan-degrading exo- and endo-enzymes. These exo- and endo-enzymes are highly effective in reducing biofilm biomass without killing microorganisms, as well as alleviating the production of an acidic pH environment conducive to tooth decay. To corroborate these results, we present biophysical evidence using single-molecule atomic force microscopy, biofilm shearing, and enamel surface topography analyses. Data show a drastic decrease in binding forces of GtfB to C. albicans (∼15-fold reduction) following enzyme treatment. Furthermore, enzymatic activity disrupted biofilm mechanical stability and significantly reduced human tooth enamel demineralization without cytotoxic effects on gingival keratinocytes. Our results represent significant progress toward a novel nonbiocidal therapeutic intervention against pathogenic bacterial-fungal biofilms by targeting the interkingdom receptor-ligand binding interactions. IMPORTANCE Biofilm formation is a key virulence factor responsible for various infectious diseases. Particularly, interactions between a fungus, Candida albicans, and a bacterium, Streptococcus mutans, have been known to play important roles in the pathogenesis of dental caries. Although some antimicrobials have been applied to treat fungal-involved biofilm-associated diseases, these often lack targeting polymicrobial interactions. Furthermore, these may not be appropriate for preventive measures because these antimicrobials may disrupt ecological microbiota and/or induce the prevalence of drug resistance over time. By specifically targeting the interaction mechanism whereby mannoproteins on the C. albicans surface mediate the cross-kingdom interaction, we demonstrated that mannoprotein-degrading enzymes can effectively disrupt biofilm interactions without microbiocidal effects or causing cytotoxicity to human cells. This suggests a potential application as a targeted approach for intervening a pathogenic cross-kingdom biofilm associated with a costly and unresolved oral disease.

scholarly journals Role of Glucosyltransferase B in Interactions of Candida albicans with Streptococcus mutans and with an Experimental Pellicle on Hydroxyapatite Surfaces

2011 ◽  
Vol 77 (18) ◽  
pp. 6357-6367 ◽  
Author(s):  
S. Gregoire ◽  
J. Xiao ◽  
B. B. Silva ◽  
I. Gonzalez ◽  
P. S. Agidi ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Cell Surface ◽  
Yeast Cell ◽  
Fluorescence Imaging ◽  
Streptococcus Mutans ◽  
Tooth Enamel ◽  
Yeast Cells ◽  
Content Type ◽  
Yeast Cell Surface

ABSTRACTCandida albicansand mutans streptococci are frequently detected in dental plaque biofilms from toddlers afflicted with early childhood caries. Glucosyltransferases (Gtfs) secreted byStreptococcus mutansbind to saliva-coated apatite (sHA) and to bacterial surfaces, synthesizing exopolymersin situ, which promote cell clustering and adherence to tooth enamel. We investigated the potential role Gtfs may play in mediating the interactions betweenC. albicansSC5314 andS. mutansUA159, both with each other and with the sHA surface. GtfB adhered effectively to theC. albicansyeast cell surface in an enzymatically active form, as determined by scintillation spectroscopy and fluorescence imaging. The glucans formed on the yeast cell surface were more susceptible to dextranase than those synthesized in solution or on sHA and bacterial cell surfaces (P< 0.05), indicating an elevated α-1,6-linked glucose content. Fluorescence imaging revealed that larger numbers ofS. mutanscells bound toC. albicanscells with glucans present on their surface than to yeast cells without surface glucans (uncoated). The glucans formedin situalso enhancedC. albicansinteractions with sHA, as determined by a novel single-cell micromechanical method. Furthermore, the presence of glucan-coated yeast cells significantly increased the accumulation ofS. mutanson the sHA surface (versusS. mutansincubated alone or mixed with uncoatedC. albicans;P< 0.05). These data reveal a novel cross-kingdom interaction that is mediated by bacterial GtfB, which readily attaches to the yeast cell surface. Surface-bound GtfB promotes the formation of a glucan-rich matrixin situand may enhance the accumulation ofS. mutanson the tooth enamel surface, thereby modulating the development of virulent biofilms.

scholarly journals Multi-omics Analyses Reveal Synergistic Carbohydrate Metabolism in Streptococcus mutans-Candida albicans Mixed-Species Biofilms

2019 ◽  
Vol 87 (10) ◽  
Author(s):  
K. Ellepola ◽  
T. Truong ◽  
Y. Liu ◽  
Q. Lin ◽  
T. K. Lim ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Carbohydrate Metabolism ◽  
Streptococcus Mutans ◽  
Quantitative Proteomics ◽  
Sugar Transport ◽  
Oral Disease ◽  
Mixed Species ◽  
Carbohydrate Utilization ◽  
Content Type ◽  
The Impact

ABSTRACT Candida albicans, a major opportunistic fungal pathogen, is frequently found together with Streptococcus mutans in dental biofilms associated with severe childhood caries (tooth decay), a prevalent pediatric oral disease. However, the impact of this cross-kingdom relationship on C. albicans remains largely uncharacterized. Here, we employed a novel quantitative proteomics approach in conjunction with transcriptomic profiling to unravel molecular pathways of C. albicans when cocultured with S. mutans in mixed biofilms. RNA sequencing and iTRAQ (isobaric tags for relative and absolute quantitation)-based quantitative proteomics revealed that C. albicans genes and proteins associated with carbohydrate metabolism were significantly enhanced, including sugar transport, aerobic respiration, pyruvate breakdown, and the glyoxylate cycle. Other C. albicans genes and proteins directly and indirectly related to cell morphogenesis and cell wall components such as mannan and glucan were also upregulated, indicating enhanced fungal activity in mixed-species biofilm. Further analyses revealed that S. mutans-derived exoenzyme glucosyltransferase B (GtfB), which binds to the fungal cell surface to promote coadhesion, can break down sucrose into glucose and fructose that can be readily metabolized by C. albicans, enhancing growth and acid production. Altogether, we identified key pathways used by C. albicans in the mixed biofilm, indicating an active fungal role in the sugar metabolism and environmental acidification (key virulence traits associated with caries onset) when interacting with S. mutans, and a new cross-feeding mechanism mediated by GtfB that enhances C. albicans carbohydrate utilization. In addition, we demonstrate that comprehensive transcriptomics and quantitative proteomics can be powerful tools to study microbial contributions which remain underexplored in cross-kingdom biofilms.

scholarly journals Understanding the Basis of METH Mouth Using a Rodent Model of Methamphetamine Injection, Sugar Consumption, and Streptococcus mutans Infection

mBio ◽  
2021 ◽  
Vol 12 (2) ◽  
Author(s):  
Hiu Ham Lee ◽  
Preethi Sudhakara ◽  
Shreena Desai ◽  
Kildare Miranda ◽  
Luis R. Martinez
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Human Saliva ◽  
Tooth Decay ◽  
Content Type ◽  
Oral Rinse ◽  
Tooth Grinding ◽  
Oral Tissue ◽  
Public Health Strategies ◽  
The Impact

ABSTRACT “METH mouth” is a common consequence of chronic methamphetamine (METH) use, resulting in tooth decay and painful oral tissue inflammation that can progress to complete tooth loss. METH reduces the amount of saliva in the mouth, promoting bacterial growth, tooth decay, and oral tissue damage. This oral condition is worsened by METH users’ compulsive behavior, including high rates of consumption of sugary drinks, recurrent tooth grinding, and a lack of frequent oral hygiene. Streptococcus mutans is a Gram-positive bacterium found in the oral cavity and associated with caries in humans. Hence, we developed a murine model of METH administration, sugar intake, and S. mutans infection to mimic METH mouth in humans and to investigate the impact of this drug on tooth colonization. We demonstrated that the combination of METH and sucrose stimulates S. mutans tooth adhesion, growth, and biofilm formation in vivo. METH and sucrose increased the expression of S. mutans glycosyltransferases and lactic acid production. Moreover, METH contributes to the low environmental pH and S. mutans sucrose metabolism, providing a plausible mechanism for bacterium-mediated tooth decay. Daily oral rinse treatment with chlorhexidine significantly reduces tooth colonization in METH- and sucrose-treated mice. Furthermore, human saliva inhibits S. mutans colonization and biofilm formation after exposure to either sucrose or the combination of METH and sucrose. These findings suggest that METH might increase the risk of microbial dental disease in users, information that may help in the development of effective public health strategies to deal with this scourge in our society. IMPORTANCE “METH mouth” is characterized by severe tooth decay and gum disease, which often causes teeth to break or fall out. METH users are also prone to colonization by cariogenic bacteria such as Streptococcus mutans. In addition, this oral condition is aggravated by METH users’ compulsive behavior, including the consumption of beverages with high sugar content, recurrent tooth grinding, and a lack of frequent oral hygiene. We investigated the effects of METH and sugar consumption on S. mutans biofilm formation and tooth colonization. Using a murine model of METH administration, sucrose ingestion, and oral infection, we found that the combination of METH and sucrose increases S. mutans adhesion and biofilm formation on the teeth of C57BL/6 mice. However, daily chlorhexidine-based oral rinse treatment reduces S. mutans tooth colonization. Similarly, METH has been associated with dry mouth or hyposalivation in users. Hence, we assessed the impact of human saliva on biofilm formation and demonstrated that surface preconditioning with saliva substantially reduces S. mutans biofilm formation. Our results are significant because to our knowledge, this is the first basic science study focused on elucidating the fundamentals of METH mouth using a rodent model of prolonged drug injection and S. mutans oral infection. Our findings may have important translational implications for the development of treatments for the management of METH mouth and more effective preventive public health strategies that can be applied to provide effective dental care for METH users in prisons, drug treatment centers, and health clinics.

Diagnosis and detection of VicK gene in Streptococcus mutans isolated from the saliva of patients with diabetic type 2 with tooth decay in the Iraqi population

2022 ◽  
Vol 67 (4) ◽  
pp. 222-231
Author(s):  
Susan F.Khadhem Al-Sudani ◽  
Laheeb R. Hamad ◽  
Fattma A. Ali
Keyword(s):  
Type 2 Diabetes ◽  
Streptococcus Mutans ◽  
Phylogenetic Trees ◽  
Root Surface ◽  
Oral Disease ◽  
Tooth Decay ◽  
Random Blood Sugar ◽  
Severe Periodontitis ◽  
Pcr Products

Type 2 diabetes mellitus (T2DM) is gradually becoming more common in Iraq. Salivary changes and proliferation of specific bacterial communities cause oral disease that can adversely affect systemic conditions such as diabetes. Fifty saliva samples were collected from people with T2DM suffering from tooth decay and twenty-five people without T2DM suffering from tooth decay. The periodontal status, the extent of the root surface, and coronal caries were evaluated. Saliva was cultured for investigating Streptococcus mutans. The results showed that patients with type 2 diabetes had significantly more severe Periodontitis and a higher prevalence and magnitude of bacterial caries. Diabetic subjects had higher levels of Hemoglobin A1c (HbA1c) and Random Blood Sugar (R.B.S.). The S. mutans diagnosis by PCR for Sanger Sequencing technique by using VicK gene sequences (1300bp). The PCR products of the isolate were submitted to Macrogen Company for sequencing. Selected seven isolates as new isolates registered in global gene bank as locally S. Mutans isolates in Bagdad city/Iraq and their accepted accession numbers include LOCUS MT603520, MT603521, MT603522, MT603523, MT603524, MT603525,and MT603526 of nucleotide sequence. The VicK genes isolates' phylogenetic trees revealed a genotype that was closely connected to other isolates in GenBank. Furthermore, gene sequencing demonstrated a success rate of 99 percent. resemblance to other isolates in the GenBank database The likelihood of a link between S. Mutans and dental carries was determined by these findings.

scholarly journals Metabolic Modeling of Streptococcus mutans Reveals Complex Nutrient Requirements of an Oral Pathogen

mSystems ◽  
2019 ◽  
Vol 4 (5) ◽  
Author(s):  
Kenan Jijakli ◽  
Paul A. Jensen
Keyword(s):  
Streptococcus Mutans ◽  
Single Gene ◽  
Metabolic Model ◽  
The United States ◽  
Defined Medium ◽  
Oral Microbiome ◽  
Tooth Decay ◽  
Oral Pathogen ◽  
Content Type ◽  
A Genome

ABSTRACT Streptococcus mutans is a Gram-positive bacterium that thrives under acidic conditions and is a primary cause of tooth decay (dental caries). To better understand the metabolism of S. mutans on a systematic level, we manually constructed a genome-scale metabolic model of the S. mutans type strain UA159. The model, called iSMU, contains 675 reactions involving 429 metabolites and the products of 493 genes. We validated iSMU by comparing simulations with growth experiments in defined medium. The model simulations matched experimental results for 17 of 18 carbon source utilization assays and 47 of 49 nutrient depletion assays. We also simulated the effects of single gene deletions. The model’s predictions agreed with 78.1% and 84.4% of the gene essentiality predictions from two experimental data sets. Our manually curated model is more accurate than S. mutans models generated from automated reconstruction pipelines and more complete than other manually curated models. We used iSMU to generate hypotheses about the S. mutans metabolic network. Subsequent genetic experiments confirmed that (i) S. mutans catabolizes sorbitol via a sorbitol-6-phosphate 2-dehydrogenase (SMU_308) and (ii) the Leloir pathway is required for growth on complex carbohydrates such as raffinose. We believe the iSMU model is an important resource for understanding the metabolism of S. mutans and guiding future experiments. IMPORTANCE Tooth decay is the most prevalent chronic disease in the United States. Decay is caused by the bacterium Streptococcus mutans, an oral pathogen that ferments sugars into tooth-destroying lactic acid. We constructed a complete metabolic model of S. mutans to systematically investigate how the bacterium grows. The model provides a valuable resource for understanding and targeting S. mutans’ ability to outcompete other species in the oral microbiome.

scholarly journals Symbiotic Relationship between Streptococcus mutans and Candida albicans Synergizes Virulence of Plaque BiofilmsIn Vivo

2014 ◽  
Vol 82 (5) ◽  
pp. 1968-1981 ◽  
Author(s):  
Megan L. Falsetta ◽  
Marlise I. Klein ◽  
Punsiri M. Colonne ◽  
Kathleen Scott-Anne ◽  
Stacy Gregoire ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Dental Caries ◽  
Streptococcus Mutans ◽  
Bacterial Pathogen ◽  
Single Species ◽  
Content Type ◽  
3 Dimensional ◽  
Biofilm Development

ABSTRACTStreptococcus mutansis often cited as the main bacterial pathogen in dental caries, particularly in early-childhood caries (ECC).S. mutansmay not act alone;Candida albicanscells are frequently detected along with heavy infection byS. mutansin plaque biofilms from ECC-affected children. It remains to be elucidated whether this association is involved in the enhancement of biofilm virulence. We showed that the ability of these organisms together to form biofilms is enhancedin vitroandin vivo. The presence ofC. albicansaugments the production of exopolysaccharides (EPS), such that cospecies biofilms accrue more biomass and harbor more viableS. mutanscells than single-species biofilms. The resulting 3-dimensional biofilm architecture displays sizeableS. mutansmicrocolonies surrounded by fungal cells, which are enmeshed in a dense EPS-rich matrix. Using a rodent model, we explored the implications of this cross-kingdom interaction for the pathogenesis of dental caries. Coinfected animals displayed higher levels of infection and microbial carriage within plaque biofilms than animals infected with either species alone. Furthermore, coinfection synergistically enhanced biofilm virulence, leading to aggressive onset of the disease with rampant carious lesions. Ourin vitrodata also revealed that glucosyltransferase-derived EPS is a key mediator of cospecies biofilm development and that coexistence withC. albicansinduces the expression of virulence genes inS. mutans(e.g.,gtfB,fabM). We also found thatCandida-derived β1,3-glucans contribute to the EPS matrix structure, while fungal mannan and β-glucan provide sites for GtfB binding and activity. Altogether, we demonstrate a novel mutualistic bacterium-fungus relationship that occurs at a clinically relevant site to amplify the severity of a ubiquitous infectious disease.

Lemon myrtle extract inhibits lactate production by Streptococcus mutans

2021 ◽  
Vol 85 (10) ◽  
pp. 2185-2190
Author(s):  
Yukinori Yabuta ◽  
Yui Sato ◽  
Arisu Miki ◽  
Ryuta Nagata ◽  
Tomohiro Bito ◽  
...  
Keyword(s):  
Lactate Dehydrogenase ◽  
Dental Caries ◽  
Oral Cavity ◽  
Streptococcus Mutans ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Tooth Enamel ◽  
Lactate Production ◽  
Tooth Decay ◽  
Lactate Dehydrogenase Activity

ABSTRACT Backhousia citriodora (lemon myrtle) extract has been found to inhibit glucansucrase activity, which plays an important role in biofilm formation by Streptococcus mutans. In addition to glucansucrase, various virulence factors in S. mutans are involved in the initiation of caries. Lactate produced by S. mutans demineralizes the tooth enamel. This study investigated whether lemon myrtle extract can inhibit S. mutans lactate production. Lemon myrtle extract reduced the glycolytic pH drop in S. mutans culture and inhibited lactate production by at least 46%. Ellagic acid, quercetin, hesperetin, and myricetin, major polyphenols in lemon myrtle, reduced the glycolytic pH drop and lactate production, but not lactate dehydrogenase activity. Furthermore, these polyphenols reduced the viable S. mutans cell count. Thus, lemon myrtle extracts may inhibit S. mutans-mediated acidification of the oral cavity, thereby preventing dental caries and tooth decay.

scholarly journals Streptococcus mutans yidC1andyidC2Impact Cell Envelope Biogenesis, the Biofilm Matrix, and Biofilm Biophysical Properties

2018 ◽  
Vol 201 (1) ◽  
Author(s):  
Sara R. Palmer ◽  
Zhi Ren ◽  
Geelsu Hwang ◽  
Yuan Liu ◽  
Ashton Combs ◽  
...  
Keyword(s):  
Physical Properties ◽  
Streptococcus Mutans ◽  
Spatial Organization ◽  
Mechanical Stability ◽  
Cell Envelope ◽  
Wild Type ◽  
Biophysical Properties ◽  
Content Type ◽  
Glucan Synthesis ◽  
Biofilm Development

ABSTRACTProper envelope biogenesis ofStreptococcus mutans, a biofilm-forming and dental caries-causing oral pathogen, requires two paralogs (yidC1andyidC2) of the universally conserved YidC/Oxa1/Alb3 family of membrane integral chaperones and insertases. The deletion of either paralog attenuates virulencein vivo, but the mechanisms of disruption remain unclear. Here, we determined whether the deletion ofyidCaffects cell surface properties, extracellular glucan production, and/or the structural organization of the exopolysaccharide (EPS) matrix and biophysical properties ofS. mutansbiofilm. Compared to the wild type, the ΔyidC2 mutant lacked staining with fluorescent vancomycin at the division septum, while the ΔyidC1mutant resembled the wild type. Additionally, the deletion of eitheryidC1oryidC2resulted in less insoluble glucan synthesis but produced more soluble glucans, especially at early and mid-exponential-growth phases. Alteration of glucan synthesis by both mutants yielded biofilms with less dry weight and insoluble EPS. In particular, the deletion ofyidC2resulted in a significant reduction in biofilm biomass and pronounced defects in the spatial organization of the EPS matrix, thus modifying the three-dimensional (3D) biofilm architecture. The defective biofilm harbored smaller bacterial clusters with high cell density and less surrounding EPS than those of the wild type, which was stiffer in compression yet more susceptible to removal by shear. Together, our results indicate that the elimination of eitheryidCparalog results in changes to the cell envelope and glucan production that ultimately disrupts biofilm development and EPS matrix structure/composition, thereby altering the physical properties of the biofilms and facilitating their removal. YidC proteins, therefore, represent potential therapeutic targets for cariogenic biofilm control.IMPORTANCEYidC proteins are membrane-localized chaperone insertases that are universally conserved in all bacteria and are traditionally studied in the context of membrane protein insertion and assembly. Both YidC paralogs of the cariogenic pathogenStreptococcus mutansare required for proper envelope biogenesis and full virulence, indicating that these proteins may also contribute to optimal biofilm formation in streptococci. Here, we show that the deletion of eitheryidCresults in changes to the structure and physical properties of the EPS matrix produced byS. mutans, ultimately impairing optimal biofilm development, diminishing its mechanical stability, and facilitating its removal. Importantly, the universal conservation of bacterialyidCorthologs, combined with our findings, provide a rationale for YidC as a possible drug target for antibiofilm therapies.

scholarly journals Cell Toxicity and inhibitory effects of Cyperus rotundus extract on Streptococcus mutans, Aggregatibacter actinomycetemcomitans and Candida albicans

2018 ◽  
Vol 28 (4) ◽  
Author(s):  
Moein Khojaste ◽  
Mohsen Yazdanian ◽  
Elahe Tahmasebi ◽  
Mehdi Shokri ◽  
Behzad Houshmand ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Essential Oil ◽  
Streptococcus Mutans ◽  
Antimicrobial Effect ◽  
Aggregatibacter Actinomycetemcomitans ◽  
Cyperus Rotundus ◽  
Tooth Decay ◽  
Gingival Fibroblast ◽  
Alcoholic Extract ◽  
Aqueous Extracts

Periodontitis and tooth decay are common dental diseases. There are some bacterial risk factors such as Streptococcus mutans, Aggregatibacter actinomycetemcomitans and Candida albicans complications, tubers of Cyperus rotundus was used to determine the antimicrobial effect of fungi. Considering the increasing resistance of bacteria and fungi to antibiotics and their alcoholic and aqueous extracts and essential oil on Streptococcus mutans, Aggregatibacter actinomycetemcomitans and Candida albicans, as well as an examination cytotoxicity on gingival fibroblast cells. Alcoholic and aqueous extracts and essential oil prepared from tubers of Cyperus rotundus. After bacterial and fungal cultures, the inhibition zone, Minimum inhibitory concentrations (MIC), and Minimum Bactericidal Concentration (MBC), were studied. Cytotoxicity was also evaluated using the Methyl tetrazolium test (MTT).The results showed that Cyperus rotundus alcoholic extract had the greatest effect on inhibition of growth and death of Streptococcus mutans, Aggregatibacter actinomycetemcomitans. According to the present study, Cyperus rotundus extract can be considered as a suitable candidate for the treatment and prevention of periodontitis and tooth decay.

Sign in / Sign up

Export Citation Format

Share Document