scholarly journals Genetic Analysis of Mutacin B-Ny266, a Lantibiotic Active against Caries Pathogens

2020 ◽  
Vol 202 (12) ◽  
Author(s):  
Delphine Dufour ◽  
Abdelahhad Barbour ◽  
Yuki Chan ◽  
Marcus Cheng ◽  
Taimoor Rahman ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Dental Caries ◽  
Streptococcus Mutans ◽  
Dental Plaque ◽  
Oral Bacteria ◽  
Oral Streptococci ◽  
Chromosomal Locus ◽  
Content Type ◽  
Genes Encoding ◽  
Plaque Biofilm

ABSTRACT Bacteriocins are ribosomally synthesized proteinaceous antibacterial peptides. They selectively interfere with the growth of other bacteria. The production and secretion of bacteriocins confer a distinct ecological advantage to the producer in competing against other bacteria that are present in the same ecological niche. Streptococcus mutans, a significant contributor to the development of dental caries, is one of the most prolific producers of bacteriocins, known as mutacins in S. mutans. In this study, we characterized the locus encoding mutacin B-Ny266, a lantibiotic with a broad spectrum of activity. The chromosomal locus is composed of six predicted operon structures encoding proteins involved in regulation, antimicrobial activity, biosynthesis, modification, transport, and immunity. Mutacin B-Ny266 was purified from semisolid cultures, and two inhibitory peptides, LanA and LanA′, were detected. Both peptides were highly modified. Such modifications include dehydration of serine and threonine and the formation of a C-terminal aminovinyl-cysteine (AviCys) ring. While LanA peptide alone is absolutely required for antimicrobial activity, the presence of LanA′ enhanced the activity of LanA, suggesting that B-Ny266 may function as a two-peptide lantibiotic. The activation of lanAA′ expression is most likely controlled by the conserved two-component system NsrRS, which is activated by LanA peptide but not by LanA′. The chromosomal locus encoding mutacin B-Ny266 was not universally conserved in all sequenced S. mutans genomes. Intriguingly, the genes encoding LanAA′ peptides were restricted to the most invasive serotypes of S. mutans. IMPORTANCE Although dental caries is largely preventable, it remains the most common and costly infectious disease worldwide. Caries is initiated by the presence of dental plaque biofilm that contains Streptococcus mutans, a species extensively characterized by its role in caries development and formation. S. mutans deploys an arsenal of strategies to establish itself within the oral cavity. One of them is the production of bacteriocins that confer a competitive advantage by targeting and killing closely related competitors. In this work, we found that mutacin B-Ny266 is a potent lantibiotic that is effective at killing a wide array of oral streptococci, including nearly all S. mutans strains tested. Lantibiotics produced by oral bacteria could represent a promising strategy to target caries pathogens embedded in dental plaque biofilm.

scholarly journals Isolation and characterization of Streptococcus mutans in heart valve and dental plaque specimens from a patient with infective endocarditis

2006 ◽  
Vol 55 (8) ◽  
pp. 1135-1140 ◽  
Author(s):  
Ryota Nomura ◽  
Kazuhiko Nakano ◽  
Hirotoshi Nemoto ◽  
Kazuyo Fujita ◽  
Satoko Inagaki ◽  
...  
Keyword(s):  
Infective Endocarditis ◽  
Dental Caries ◽  
Oral Cavity ◽  
Streptococcus Mutans ◽  
Heart Valve ◽  
Dental Plaque ◽  
Biochemical Properties ◽  
Oral Streptococci ◽  
Isolation And Characterization

Streptococcus mutans, known to be an aetiologic agent of dental caries, also causes infective endocarditis (IE), although a comparison of isolates from the oral cavity and infected heart valve of the same patient has not been reported. In the present study, infected heart valve and dental plaque samples from a patient with IE were analysed. Broad-range PCR with DNA sequencing revealed that 50 clones from the dental plaque isolates were composed of oral streptococci and periodontopathic bacteria, whereas only Streptococcus mutans was detected in 50 clones from the heart valve. Eighteen strains of Streptococcus mutans were isolated from dental plaque and seven from the heart valve, and the biochemical properties of each were in accordance with those of Streptococcus mutans. DNA fingerprinting analysis revealed that all the oral isolates of Streptococcus mutans had similar patterns, which were different from those of the isolates from the infected heart valve. Western blotting using glucosyltransferase (GTF)-specific antiserum showed that the seven strains from the heart valve lacked the three types of intact GTF. In addition, the sucrose-dependent adhesion rates of these isolates were significantly lower than those of the oral isolates (P<0.001). Furthermore, the isolates from the heart valve were less susceptible to erythromycin and kanamycin. These results indicate that the properties of the Streptococcus mutans strains isolated from the infected valve were different from those of typical oral strains, which may be related to the effects of IE.

scholarly journals Contribution of Severe Dental Caries Induced by Streptococcus mutans to the Pathogenicity of Infective Endocarditis

2020 ◽  
Vol 88 (7) ◽  
Author(s):  
Ryota Nomura ◽  
Saaya Matayoshi ◽  
Masatoshi Otsugu ◽  
Takahiro Kitamura ◽  
Noboru Teramoto ◽  
...  
Keyword(s):  
Infective Endocarditis ◽  
Dental Caries ◽  
Streptococcus Mutans ◽  
Heart Tissue ◽  
Oral Bacteria ◽  
Sequencing Analysis ◽  
Dependent Manner ◽  
Content Type ◽  
Number Of Teeth ◽  
Heart Injury

ABSTRACT Streptococcus mutans, a major pathogen of dental caries, is regarded as a causative agent of infective endocarditis (IE), which mainly occurs in patients with underlying heart disease. However, it remains unknown whether severe dental caries that extend to pulp space represent a possible route of infection. In the present study, we evaluated the virulence of S. mutans for IE development using rats with concurrent severe dental caries and heart valve injury. Dental caries was induced in rats through the combination of a caries-inducing diet and the administration of S. mutans into the oral cavity. Then, the heart valves of a subset of rats were injured using a sterile catheter and wire under general anesthesia. The rats were euthanized at various times with various stages of dental caries. The number of teeth affected by dental caries with pulp exposure was increased in the rats in a time-dependent manner. S. mutans was recovered from injured heart tissue, which was mainly observed in rats with higher number of S. mutans bacteria in mandibular bone and a larger number of teeth in which caries extended to pulp. Dental caries was more severe in rats with heart injury than in rats without heart injury. Sequencing analysis targeting 16S rRNA revealed that specific oral bacteria appeared only in rats with heart injury, which may be related to the development of dental caries. Our findings suggest that dental caries caused by the combination of S. mutans infection and sucrose intake may contribute to S. mutans colonization in injured heart tissue.

ACTIVITY OF PERICARP EXTRACT OF MANGOSTEEN AGAINST ORAL STREPTOCOCCI

2017 ◽  
Vol 20 (1) ◽  
pp. 41-46
Author(s):  
Dewi Nurul Mustaqimah ◽  
Josh Erry HW
Keyword(s):  
Antimicrobial Activity ◽  
Dental Caries ◽  
Streptococcus Mutans ◽  
Antimicrobial Agents ◽  
Biological Activities ◽  
World Health ◽  
Direct Result ◽  
Oral Streptococci ◽  
Oral Pathogens

The increasing prevalence of dental caries is still as a major world health problem. Caries is the direct result of acid production by cariogenic oral pathogens, especially Streptococcus mutans. New and better antimicrobial agents active against cariogenic bacteria with minimal side effects on the oral tissues are much needed, especially natural agents derived directly from plants. Phytochemical studies have shown that the extracts from various parts of mangosteen or Garciniamangostana Linn tree contain varieties of secondary metabolites such as prenylated and oxygenated xanthones, many of which have been found in vitro to have antimicrobial properties against oral pathogens. Several studies which examined the eficacy of herbal for human health have shown that xanthones from mangosteen have remarkable biological activities such as antioxidant, antimicrobial, anticancer etc, and had no cytotoxic effects on human gingival fibroblasts. Their results showed that among these xanthone derivatives obtain from pericarp extract of mangosteen, α-mangostin has the most potent antimicrobial activity against cariogenic Streptococcus mutans. It can be concluded that the strong antimicrobial activity of the pericarp extract of mangosteen is a good drug of choice that might be helpful in preventing the dental caries.

scholarly journals Amino Sugars Enhance the Competitiveness of Beneficial Commensals with Streptococcus mutans through Multiple Mechanisms

2016 ◽  
Vol 82 (12) ◽  
pp. 3671-3682 ◽  
Author(s):  
Lin Zeng ◽  
Tanaz Farivar ◽  
Robert A. Burne
Keyword(s):  
Dental Caries ◽  
Streptococcus Mutans ◽  
Sugar Metabolism ◽  
Amino Sugar ◽  
Amino Sugars ◽  
Mrna Levels ◽  
Oral Streptococci ◽  
Streptococcus Gordonii ◽  
Carbohydrate Source ◽  
Content Type

ABSTRACTBiochemical and genetic aspects of the metabolism of the amino sugarsN-acetylglucosamine (GlcNAc) and glucosamine (GlcN) by commensal oral streptococci and the effects of these sugars on interspecies competition with the dental caries pathogenStreptococcus mutanswere explored. MultipleS. mutanswild-type isolates displayed long lag phases when transferred from glucose-containing medium to medium with GlcNAc as the primary carbohydrate source, but commensal streptococci did not. Competition in liquid coculture or dual-species biofilms betweenS. mutansandStreptococcus gordoniishowed thatS. gordoniiwas particularly dominant when the primary carbohydrate was GlcN or GlcNAc. Transcriptional and enzymatic assays showed that the catabolic pathway for GlcNAc was less highly induced inS. mutansthan inS. gordonii. Exposure to H2O2, which is produced byS. gordoniiand antagonizes the growth ofS. mutans, led to reduced mRNA levels ofnagAandnagBinS. mutans. When the gene for the transcriptional regulatory NagR was deleted inS. gordonii, the strain produced constitutively high levels ofnagA(GlcNAc-6-P deacetylase),nagB(GlcN-6-P deaminase), andglmS(GlcN-6-P synthase) mRNA. Similar to NagR ofS. mutans(NagRSm), theS. gordoniiNagR protein (NagRSg) could bind to consensus binding sites (dre) in thenagA,nagB, andglmSpromoter regions ofS. gordonii. Notably, NagRSgbinding was inhibited by GlcN-6-P, but G-6-P had no effect, unlike for NagRSm. This study expands the understanding of amino sugar metabolism and NagR-dependent gene regulation in streptococci and highlights the potential for therapeutic applications of amino sugars to prevent dental caries.IMPORTANCEAmino sugars are abundant in the biosphere, so the relative efficiency of particular bacteria in a given microbiota to metabolize these sources of carbon and nitrogen might have a profound impact on the ecology of the community. Our investigation reveals that several oral commensal bacteria have a much greater capacity to utilize amino sugars than the dental pathogenStreptococcus mutansand that the ability of the model commensalStreptococcus gordoniito compete againstS. mutansis substantively enhanced by the presence of amino sugars commonly found in the oral cavity. The mechanisms underlying the greater capacity and competitive enhancements of the commensal are shown to depend on how the genes for the catabolic enzymes are regulated, the role of the allosteric modulators affecting such regulation, and the ability of amino sugars to enhance certain activities of the commensal that are antagonistic toS. mutans.

scholarly journals Antimicrobial Activity of Silver Nanoparticles against Clinical Biofilms from Patients with and without Dental Caries

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Alondra Jaqueline Jiménez-Ramírez ◽  
Rita Elizabeth Martínez-Martínez ◽  
José Luis Ayala-Herrera ◽  
Erasto Armando Zaragoza-Contreras ◽  
Rubén Abraham Domínguez-Pérez ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Silver Nanoparticles ◽  
Dental Caries ◽  
Dental Plaque ◽  
Scientific Information ◽  
Antimicrobial Properties ◽  
Inhibitory Effect ◽  
Oral Bacteria ◽  
Oral Disease ◽  
Microbiological Analysis

Dental caries is an infectious oral disease originated by the presence of different microorganisms from well-defined biofilms. Many treatments for dental caries have been demonstrated to be successful protocols; however, incidence and prevalence remain still high. Although silver nanoparticles (AgNPs) have shown excellent antimicrobial properties, even against different oral bacteria, there is no available scientific information that has evaluated the antimicrobial activity of AgNPs against oral biofilms from subjects with active dental caries. The objective of this research was to evaluate the inhibitory effect of AgNPs in dental biofilms from subjects with and without dental caries. Two sizes of AgNPs were prepared and characterized. Dental plaque samples were collected from 30 subjects with dental caries and 30 subjects with no dental caries. Microbiological analysis was determined by the minimum inhibitory concentration (MIC) of nanoparticles, and the presence and distribution of microbial strains were identified by polymerase chain reaction (PCR) assay. AgNPs had significant antimicrobial effects against all samples of dental plaque; however, the physical properties of AgNPs, as well as specific sociodemographic and clinical conditions from patients, were also associated with bacterial growth inhibition of Ag particles ( p < 0.05 ). PCR confirmed the presence of oral bacteria associated with dental caries, such as S. mutans and S. sobrinus strains, principally. The AgNPs exhibited great potential to be used as an antimicrobial therapy for the control and prevention of dental caries.

scholarly journals Differential Utilization of Basic Proline-Rich Glycoproteins during Growth of Oral Bacteria in Saliva

2016 ◽  
Vol 82 (17) ◽  
pp. 5249-5258 ◽  
Author(s):  
Yuan Zhou ◽  
Jinghua Yang ◽  
Luxia Zhang ◽  
Xuedong Zhou ◽  
John O. Cisar ◽  
...  
Keyword(s):  
Dental Plaque ◽  
Oral Bacteria ◽  
Intact Protein ◽  
Major Protein ◽  
Streptococcus Sobrinus ◽  
Two Dimensional Gel Electrophoresis ◽  
Content Type ◽  
Whole Saliva ◽  
Biofilm Development ◽  
Plaque Biofilm

ABSTRACTAlthough saliva is widely recognized as a primary source of carbon and nitrogen for growth of the dental plaque biofilm community, little is known about how different oral bacteria utilize specific salivary components. To address this question, 32 strains representing 16 genera commonly isolated from early plaque biofilms were compared for growth over two transfers in stimulated (by chewing Parafilm) whole saliva that was stabilized by heat treatment and dialysis. The cell densities, measured by quantitative PCR (qPCR), ranged from ∼1 × 106to 1 × 107/ml for strains ofStreptococcus gordonii,Streptococcus oralis, andStreptococcus mitisand one strain ofStreptococcus sanguinis. Strains ofStreptococcus mutans,Gemella haemolysans, andGranulicatella adiacensreached ∼1 × 105to 1 × 106/ml. In contrast, little or no growth was noted for three other strains ofS. sanguinis, as well as for strains ofStreptococcus parasanguinis,Streptococcus salivarius,Streptococcus vestibularis,Streptococcus sobrinus,Actinomycesspp.,Abiotrophia defectiva, andRothia dentocariosa. SDS-PAGE, lectin blotting, and two-dimensional gel electrophoresis of saliva from cultures ofS. gordonii,S. oralis, andS. mitisrevealed species-specific differences in the degradation of basic proline-rich glycoproteins (PRG). In contrast, saliva from cultures of other bacteria was indistinguishable from control saliva. Species-dependent differences in the utilization of individual host sugars were minor. Thus, differences in salivary glycan foraging between oral species may be important to cross-feeding and cooperation between organisms in dental plaque biofilm development.IMPORTANCEBacteria in the mouth use saliva for nutrition. How each of the many types of bacteria uses saliva is not clear. We show that a major protein in saliva, called PRG, is an important nutrition source for certain bacteria but not for others. PRG has many sugar molecules linked in chains, but the sugar is not available for bacteria until the chains are degraded. The bacteria that can grow by digesting this protein break the sugar chains into parts which not only support their own growth but could also be available to support the growth of those bacteria that cannot use the intact protein.

scholarly journals Characterization of the Streptococcus mutans SMU.1703c-SMU.1702c Operon Reveals Its Role in Riboflavin Import and Response to Acid Stress

2020 ◽  
Vol 203 (2) ◽  
pp. e00293-20
Author(s):  
Matthew E. Turner ◽  
Khanh Huynh ◽  
Ronan K. Carroll ◽  
Sang-Joon Ahn ◽  
Kelly C. Rice
Keyword(s):  
Oral Cavity ◽  
Streptococcus Mutans ◽  
Tooth Enamel ◽  
Acid Stress ◽  
Oral Bacteria ◽  
Flavin Mononucleotide ◽  
Dependent Manner ◽  
Oral Streptococci ◽  
Uncharacterized Protein ◽  
Content Type

ABSTRACTStreptococcus mutans utilizes numerous metabolite transporters to obtain essential nutrients in the “feast or famine” environment of the human mouth. S. mutans and most other streptococci are considered auxotrophic for several essential vitamins including riboflavin (vitamin B2), which is used to generate key cofactors and to perform numerous cellular redox reactions. Despite the well-known contributions of this vitamin to central metabolism, little is known about how S. mutans obtains and metabolizes B2. The uncharacterized protein SMU.1703c displays high sequence homology to the riboflavin transporter RibU. Deletion of SMU.1703c hindered S. mutans growth in complex and defined medium in the absence of saturating levels of exogenous riboflavin, whereas deletion of cotranscribed SMU.1702c alone had no apparent effect on growth. Expression of SMU.1703c in a Bacillus subtilis riboflavin auxotroph functionally complemented growth in nonsaturating riboflavin conditions. S. mutans was also able to grow on flavin adenine dinucleotide (FAD) or flavin mononucleotide (FMN) in an SMU.1703c-dependent manner. Deletion of SMU.1703c and/or SMU.1702c impacted S. mutans acid stress tolerance, as all mutants showed improved growth at pH 5.5 compared to that of the wild type when medium was supplemented with saturating riboflavin. Cooccurrence of SMU.1703c and SMU.1702c, a hypothetical PAP2 family acid phosphatase gene, appears unique to the streptococci and may suggest a connection of SMU.1702c to the acquisition or metabolism of flavins within this genus. Identification of SMU.1703c as a RibU-like riboflavin transporter furthers our understanding of how S. mutans acquires essential micronutrients within the oral cavity and how this pathogen successfully competes within nutrient-starved oral biofilms.IMPORTANCE Dental caries form when acid produced by oral bacteria erodes tooth enamel. This process is driven by the fermentative metabolism of cariogenic bacteria, most notably Streptococcus mutans. Nutrient acquisition is key in the competitive oral cavity, and many organisms have evolved various strategies to procure carbon sources or necessary biomolecules. B vitamins, such as riboflavin, which many oral streptococci must scavenge from the oral environment, are necessary for survival within the competitive oral cavity. However, the primary mechanism and proteins involved in this process remain uncharacterized. This study is important because it identifies a key step in S. mutans riboflavin acquisition and cofactor generation, which may enable the development of novel anticaries treatment strategies via selective targeting of metabolite transporters.

scholarly journals Effects of Arginine on Streptococcus mutans Growth, Virulence Gene Expression, and Stress Tolerance

2017 ◽  
Vol 83 (15) ◽  
Author(s):  
Brinta Chakraborty ◽  
Robert A. Burne
Keyword(s):  
Dental Caries ◽  
Streptococcus Mutans ◽  
Environmental Stresses ◽  
Arginine Deiminase ◽  
Pathogenic Potential ◽  
Content Type ◽  
Chemically Defined Media ◽  
Defined Media ◽  
Oral Biofilms ◽  
Genes Encoding

ABSTRACT Streptococcus mutans is a common constituent of oral biofilms and a primary etiologic agent of human dental caries. The bacteria associated with dental caries have potent abilities to produce organic acids from dietary carbohydrates and to grow and metabolize in acidic conditions. By contrast, many commensal bacteria produce ammonia through the arginine deiminase system (ADS), which moderates the pH of oral biofilms. Arginine metabolism by the ADS is a significant deterrent to the initiation and progression of dental caries. In this study, we observed how exogenously provided l-arginine affects the growth, the virulence properties, and the tolerance of environmental stresses of S. mutans. Supplementation with 1.5% arginine (final concentration) had an inhibitory effect on the growth of S. mutans in complex and chemically defined media, particularly when cells were exposed to acid or oxidative stress. The genes encoding virulence factors required for attachment/accumulation (gtfB and spaP), bacteriocins (nlmA, nlmB, nlmD, and cipB), and the sigma factor required for competence development (comX) were downregulated during growth with 1.5% arginine. Deep sequencing of RNA (RNA-Seq) comparing the transcriptomes of S. mutans growing in chemically defined media with and without 1.5% arginine revealed differential expression of genes encoding ATP-binding cassette transporters, metal transporters, and constituents required for survival, metabolism, and biofilm formation. Therefore, the mechanisms of action by which arginine inhibits dental caries include direct adverse effects on multiple virulence-related properties of the most common human dental caries pathogen. IMPORTANCE Metabolism of the amino acid arginine by the arginine deiminase system (ADS) of certain oral bacteria raises the pH of dental plaque and provides a selective advantage to health-associated bacteria, thereby protecting the host from dental caries (cavities). Here, we examine the effects of arginine on the cavity-causing bacterium Streptococcus mutans. We find that arginine negatively impacts the growth, the pathogenic potential, and the tolerance of environmental stresses in a way that is likely to compromise the ability of S. mutans to cause disease. Using genetic and genomic techniques, multiple mechanisms by which arginine exerts its influence on virulence-related properties of S. mutans are discovered. This report demonstrates that a primary mechanism of action by which arginine inhibits the initiation and progression of dental caries may be by reducing the pathogenic potential of S. mutans.

scholarly journals Serotype distribution of Streptococcus mutans a pathogen of dental caries in cardiovascular specimens from Japanese patients

2007 ◽  
Vol 56 (4) ◽  
pp. 551-556 ◽  
Author(s):  
Kazuhiko Nakano ◽  
Hirotoshi Nemoto ◽  
Ryota Nomura ◽  
Hiromi Homma ◽  
Hideo Yoshioka ◽  
...  
Keyword(s):  
Cardiovascular Disease ◽  
Dental Caries ◽  
Streptococcus Mutans ◽  
Heart Valve ◽  
Dental Plaque ◽  
Healthy Subjects ◽  
Oral Bacteria ◽  
Atheromatous Plaque ◽  
Healthy Children ◽  
Serotype Distribution

The involvement of oral bacteria in the pathogenesis of cardiovascular disease has been studied, with Streptococcus mutans, a pathogen of dental caries, detected in cardiovascular lesions at a high frequency. However, no information is available regarding the properties of S. mutans detected in those lesions. Heart valve specimens were collected from 52 patients and atheromatous plaque specimens from 50 patients, all of whom underwent cardiovascular operations, and dental plaque specimens were taken from 41 of those subjects prior to surgery. Furthermore, saliva samples were taken from 73 sets of healthy mothers (n=73) and their healthy children (n=78). Bacterial DNA was extracted from all specimens, then analysed by PCR with S. mutans-specific and serotype-specific primer sets. The detection rates of S. mutans in the heart valve and atheromatous plaque specimens were 63 and 64 %, respectively. Non-c serotypes were identified with a significantly higher frequency in both cardiovascular and dental plaque samples from the subjects who underwent surgery as compared to serotype c, which was detected in 70–75 % of the samples from the healthy subjects. The serotype distribution in cardiovascular patients was significantly different from that in healthy subjects, suggesting that S. mutans serotype may be related to cardiovascular disease.

scholarly journals A Novel Small Molecule, ZY354, Inhibits Dental Caries-Associated Oral Biofilms

2019 ◽  
Vol 63 (5) ◽  
Author(s):  
Chenzi Zhang ◽  
Xinyi Kuang ◽  
Yuanzheng Zhou ◽  
Xian Peng ◽  
Qiang Guo ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Dental Caries ◽  
Small Molecule ◽  
Laser Scanning Microscopy ◽  
Oral Streptococci ◽  
Biofilm Inhibition ◽  
Content Type ◽  
Small Molecule Compound ◽  
Low Cytotoxicity ◽  
Multispecies Biofilms

ABSTRACT Biofilm control is a critical approach to the better management of dental caries. Antimicrobial small molecules have shown their potential in the disruption of oral biofilm and control of dental caries. The objectives of this study were to examine the antimicrobial activity and cytotoxicity of a newly designed small-molecule compound, ZY354. ZY354 was synthesized, and its cytotoxicity was evaluated in human oral keratinocytes (HOK), human gingival epithelial cells (HGE), and macrophages (RAW) by CCK-8 assays. Minimal inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs), minimum biofilm inhibition concentrations (MBICs), and minimum biofilm reduction concentrations (MBRCs) of ZY354 against common oral streptococci (i.e., Streptococcus mutans, Streptococcus gordonii, and Streptococcus sanguinis) were determined by microdilution method. The exopolysaccharide (EPS)/bacterium ratio and the dead/live bacterium ratio in the ZY354-treated multispecies biofilms were determined by confocal laser scanning microscopy, and the microbial composition was visualized and quantified by fluorescent in situ hybridization and quantitative PCR (qPCR). The demineralizing activity of ZY354-treated biofilms was evaluated by transverse microradiography. The results showed that ZY354 exhibited low cytotoxicity in HOK, HGE, and RAW cells and exhibited potent antimicrobial activity against common oral streptococci. The EPS and the abundance of S. mutans were significantly reduced after ZY354 treatment, along with an increased dead/live microbial ratio in multispecies biofilms compared to the level with the nontreated control. The ZY354-treated multispecies biofilms exhibited reduced demineralizing activity at the biofilm/enamel interface. In conclusion, the small-molecule compound ZY354 exhibits low cytotoxicity and remarkable antimicrobial activity against oral streptococci, and it may have a great potential in anticaries clinical applications.

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