Streptococcal Antagonism in Oral Biofilms: Streptococcus sanguinis and Streptococcus gordonii Interference with Streptococcus mutans

ABSTRACT Biofilms are polymicrobial, with diverse bacterial species competing for limited space and nutrients. Under healthy conditions, the different species in biofilms maintain an ecological balance. This balance can be disturbed by environmental factors and interspecies interactions. These perturbations can enable dominant growth of certain species, leading to disease. To model clinically relevant interspecies antagonism, we studied three well-characterized and closely related oral species, Streptococcus gordonii, Streptococcus sanguinis, and cariogenic Streptococcus mutans. S. sanguinis and S. gordonii used oxygen availability and the differential production of hydrogen peroxide (H2O2) to compete effectively against S. mutans. Interspecies antagonism was influenced by glucose with reduced production of H2O2. Furthermore, aerobic conditions stimulated the competence system and the expression of the bacteriocin mutacin IV of S. mutans, as well as the H2O2-dependent release of heterologous DNA from mixed cultures of S. sanguinis and S. gordonii. These data provide new insights into ecological factors that determine the outcome of competition between pioneer colonizing oral streptococci and the survival mechanisms of S. mutans in the oral biofilm.

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Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci andStreptococcus mutans

10.1101/551168 ◽

2019 ◽

Cited By ~ 1

Author(s):

Lulu Chen ◽

Brinta Chakraborty ◽

Jing Zou ◽

Robert A. Burne ◽

Lin Zeng

Keyword(s):

Streptococcus Mutans ◽

Ex Vivo ◽

Arginine Deiminase ◽

Clinical Isolates ◽

Amino Sugars ◽

Oral Streptococci ◽

Streptococcus Gordonii ◽

Oral Biofilms ◽

Low Passage

ABSTRACTN-acetylglucosamine (GlcNAc) and glucosamine (GlcN) enhance the competitiveness of the laboratory strain DL1 ofStreptococcus gordoniiagainst the caries pathogenStreptococcus mutans. Here we examine how amino sugars affect the interaction of five low-passage clinical isolates of abundant commensal streptococci withS. mutansutilizing a dual-species biofilm model. Compared to glucose, growth on GlcN or GlcNAc significantly reduced the viability ofS. mutansin co-cultures with most commensals, shifting the proportions of species. Consistent with these results, production of H2O2was increased in most commensals when growing on amino sugars, and inhibition ofS. mutansbyStreptococcus cristatus, Streptococcus oralis,orS. gordoniiwas enhanced by amino sugars on agar plates. All commensals exceptS. oralishad higher arginine deiminase activities when grown on GlcN, and in some cases GlcNAc. Inex vivobiofilms formed using pooled cell-containing saliva (CCS), the proportions ofS. mutanswere drastically diminished when GlcNAc was the primary carbohydrate. Increased production of H2O2could account in large part for the inhibitory effects of CCS biofilms. Surprisingly, amino sugars appeared to improve mutacin production byS. mutanson agar plates, suggesting that the commensals have mechanisms to actively subvert antagonism byS. mutansin co-cultures. Collectively, these findings demonstrate that amino sugars can enhance the beneficial properties of low-passage commensal oral streptococci and highlight their potential for moderating the cariogenicity of oral biofilms.SIGNIFICANCEDental caries is driven by dysbiosis of oral biofilms in which dominance by acid-producing and acid-tolerant bacteria results in loss of tooth mineral. Our previous work demonstrated the beneficial effects of amino sugars, GlcNAc and GlcN, in promoting the antagonistic properties of a health-associated oral bacterium,Streptococcus gordonii,in competition with the major caries pathogenStreptococcus mutans.Here we investigated 5 low-passage clinical isolates of the most common streptococcal species to establish how amino sugars may influence the ecology and virulence of oral biofilms. Using multiplein vitromodels, including a human saliva-derived microcosm biofilm, experiments showed significant enhancement by at least one amino sugar in the ability of most of these bacteria to suppress the caries pathogen. Therefore, our findings demonstrated the mechanism of action by which amino sugars may affect human oral biofilms to promote health.

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Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci andStreptococcus mutans

Applied and Environmental Microbiology ◽

10.1128/aem.00370-19 ◽

2019 ◽

Vol 85 (10) ◽

Cited By ~ 7

Author(s):

Lulu Chen ◽

Brinta Chakraborty ◽

Jing Zou ◽

Robert A. Burne ◽

Lin Zeng

Keyword(s):

Streptococcus Mutans ◽

Arginine Deiminase ◽

Clinical Isolates ◽

Amino Sugars ◽

Oral Streptococci ◽

Streptococcus Gordonii ◽

Content Type ◽

Oral Biofilms ◽

Passage Number ◽

Low Passage

ABSTRACTN-Acetylglucosamine (GlcNAc) and glucosamine (GlcN) enhance the competitiveness of the laboratory strain DL1 ofStreptococcus gordoniiagainst the caries pathogenStreptococcus mutans. Here, we examine how amino sugars affect the interaction of five low-passage-number clinical isolates of abundant commensal streptococci withS. mutansby utilizing a dual-species biofilm model. Compared to that for glucose, growth on GlcN or GlcNAc significantly reduced the viability ofS. mutansin cocultures with most commensals, shifting the proportions of species. Consistent with these results, production of H2O2was increased in most commensals when growing on amino sugars, and inhibition ofS. mutansbyStreptococcus cristatus,Streptococcus oralis, orS. gordoniiwas enhanced by amino sugars on agar plates. All commensals exceptS. oralishad higher arginine deiminase activities when grown on GlcN and, in some cases, GlcNAc. Inex vivobiofilms formed using pooled cell-containing saliva (CCS), the proportions ofS. mutanswere drastically diminished when GlcNAc was the primary carbohydrate. Increased production of H2O2could account in large part for the inhibitory effects of CCS biofilms. Surprisingly, amino sugars appeared to improve mutacin production byS. mutanson agar plates, suggesting that the commensals have mechanisms to actively subvert antagonism byS. mutansin cocultures. Collectively, these findings demonstrate that amino sugars can enhance the beneficial properties of low-passage-number commensal oral streptococci and highlight their potential for moderating the cariogenicity of oral biofilms.IMPORTANCEDental caries is driven by dysbiosis of oral biofilms in which dominance by acid-producing and acid-tolerant bacteria results in loss of tooth mineral. Our previous work demonstrated the beneficial effects of amino sugars GlcNAc and GlcN in promoting the antagonistic properties of a health-associated oral bacterium,Streptococcus gordonii, in competition with the major caries pathogenStreptococcus mutans. Here, we investigated 5 low-passage-number clinical isolates of the most common streptococcal species to establish how amino sugars may influence the ecology and virulence of oral biofilms. Using multiplein vitromodels, including a human saliva-derived microcosm biofilm, experiments showed significant enhancement by at least one amino sugar in the ability of most of these bacteria to suppress the caries pathogen. Therefore, our findings demonstrated the mechanism of action by which amino sugars may affect human oral biofilms to promote health.

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Competition and Coexistence between Streptococcus mutans and Streptococcus sanguinis in the Dental Biofilm

Journal of Bacteriology ◽

10.1128/jb.187.21.7193-7203.2005 ◽

2005 ◽

Vol 187 (21) ◽

pp. 7193-7203 ◽

Cited By ~ 278

Author(s):

Jens Kreth ◽

Justin Merritt ◽

Wenyuan Shi ◽

Fengxia Qi

Keyword(s):

Streptococcus Mutans ◽

Environmental Conditions ◽

Bacterial Species ◽

Interspecies Interactions ◽

Biological Factors ◽

Phenotypic Characteristics ◽

Microbial Diseases ◽

Streptococcus Sanguinis ◽

Dental Biofilm ◽

Biochemical Analyses

ABSTRACT The human mucosal surface is colonized by the indigenous microflora, which normally maintains an ecological balance among different species. Certain environmental or biological factors, however, may trigger disruption of this balance, leading to microbial diseases. In this study, we used two oral bacterial species, Streptococcus mutans and Streptococcus sanguinis (formerly S. sanguis), as a model to probe the possible mechanisms of competition/coexistence between different species which occupy the same ecological niche. We show that the two species engage in a multitude of antagonistic interactions temporally and spatially; occupation of a niche by one species precludes colonization by the other, while simultaneous colonization by both species results in coexistence. Environmental conditions, such as cell density, nutritional availability, and pH, play important roles in determining the outcome of these interactions. Genetic and biochemical analyses reveal that these interspecies interactions are possibly mediated through a well-regulated production of chemicals, such as bacteriocins (produced by S. mutans) and hydrogen peroxide (produced by S. sanguinis). Consistent with the phenotypic characteristics, production of bacteriocins and H2O2 are regulated by environmental conditions, as well as by juxtaposition of the two species. These sophisticated interspecies interactions could play an essential part in balancing competition/coexistence within multispecies microbial communities.

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Streptococcus gordonii Hsa Environmentally Constrains Competitive Binding by Streptococcus sanguinis to Saliva-Coated Hydroxyapatite

Journal of Bacteriology ◽

10.1128/jb.01535-06 ◽

2007 ◽

Vol 189 (8) ◽

pp. 3106-3114 ◽

Cited By ~ 36

Author(s):

Angela H. Nobbs ◽

Yongshu Zhang ◽

Ali Khammanivong ◽

Mark C. Herzberg

Keyword(s):

Selective Advantage ◽

Competitive Binding ◽

Tooth Surface ◽

Oral Streptococci ◽

Streptococcus Gordonii ◽

Streptococcus Sanguinis ◽

Competitive Capabilities ◽

Vitro Model ◽

Isogenic Mutants

ABSTRACT Competition between pioneer colonizing bacteria may determine polymicrobial succession during dental plaque development, but the ecological constraints are poorly understood. For example, more Streptococcus sanguinis than Streptococcus gordonii organisms are consistently isolated from the same intraoral sites, yet S. gordonii fails to be excluded and survives as a species over time. To explain this observation, we hypothesized that S. gordonii could compete with S. sanguinis to adhere to saliva-coated hydroxyapatite (sHA), an in vitro model of the tooth surface. Both species bound similarly to sHA, yet 10- to 50-fold excess S. gordonii DL1 reduced binding of S. sanguinis SK36 by 85 to >95%. S. sanguinis, by contrast, did not significantly compete with S. gordonii to adhere. S. gordonii competed with S. sanguinis more effectively than other species of oral streptococci and depended upon the salivary film on HA. Next, putative S. gordonii adhesins were analyzed for contributions to interspecies competitive binding. Like wild-type S. gordonii, isogenic mutants with mutations in antigen I/II polypeptides (sspAB), amylase-binding proteins (abpAB), and Csh adhesins (cshAB) competed effectively against S. sanguinis. By contrast, an hsa-deficient mutant of S. gordonii showed significantly reduced binding and competitive capabilities, while these properties were restored in an hsa-complemented strain. Thus, Hsa confers a selective advantage to S. gordonii over S. sanguinis in competitive binding to sHA. Hsa expression may, therefore, serve as an environmental constraint against S. sanguinis, enabling S. gordonii to persist within the oral cavity, despite the greater natural prevalence of S. sanguinis in plaque and saliva.

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In Vitro Evaluation of Bacterial Adhesion and Bacterial Viability of Streptococcus mutans, Streptococcus sanguinis, and Porphyromonas gingivalis on the Abutment Surface of Titanium and Zirconium Dental Implants

International Journal of Dentistry ◽

10.1155/2019/4292976 ◽

2019 ◽

Vol 2019 ◽

pp. 1-5 ◽

Cited By ~ 3

Author(s):

Ana Stefany Meza-Siccha ◽

Miguel Angel Aguilar-Luis ◽

Wilmer Silva-Caso ◽

Fernando Mazulis ◽

Carolina Barragan-Salazar ◽

...

Keyword(s):

Dental Implants ◽

Streptococcus Mutans ◽

Porphyromonas Gingivalis ◽

Bacterial Adhesion ◽

Bacterial Species ◽

Bacterial Adherence ◽

Bacterial Viability ◽

Streptococcus Sanguinis ◽

Group 2

Objective. To evaluate the in vitro adherence and viability of 3 bacterial species Streptococcus mutans (ATCC 25175), Streptococcus sanguinis (ATCC 10556), and Porphyromonas gingivalis (ATCC 33277) on the surfaces of dental implants of titanium, zirconium, and their respective fixing screws. Methods. Two analysis groups were formed: group 1 with 3 titanium pillars and group 2 with 3 zirconium pillars, each with their respective fixing screws. Each of these groups was included in tubes with bacterial cultures of Streptococcus mutans (ATCC 25175), Streptococcus sanguinis (ATCC 10556), and Porphyromonas gingivalis (ATCC 33277). These samples were incubated at 37°C under anaerobic conditions. Bacterial adherence was assessed by measurement of the change in colony-forming units (CFU), and bacterial viability was evaluated with the colorimetric test of 3-(4,5-dimethylthiazol-2)-2,5 diphenyl tetrazolium bromide (MTT). Results. The bacterial adhesion in the titanium abutments was higher for Streptococcus mutans (190.90 CFU/mL), and the viability was greater in Porphyromonas gingivalis (73.22%). The zirconium abutment group showed the highest adherence with Streptococcus mutans (331.82 CFU/mL) and the highest bacterial viability with the S. sanguinis strain (38.42%). The titanium fixation screws showed the highest adhesion with S. sanguinis (132.5 CFU/mL) compared to the zirconium fixation screws where S. mutans had the highest adhesion (145.5 CFU/mL). The bacterial viability of S. mutans was greater both in the titanium fixation screws and in the zirconium fixation screws 78.04% and 57.38%, respectively. Conclusions. Our results indicate that there is in vitro bacterial adherence and viability in both titanium abutments and zirconium abutments and fixation screws for both. Streptococcus mutans is the microorganism that shows the greatest adherence to the surfaces of both titanium and zirconium and the fixing screws of the latter. On the contrary, bacterial viability is greater on the titanium abutments with P. gingivalis than on the zirconium abutments with S. sanguinis. With respect to the fixation screws, in both cases, the viability of S. mutans was greater with respect to the other bacteria. In general, the titanium abutments showed less adherence but greater bacterial viability.

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

Applied and Environmental Microbiology ◽

10.1128/aem.00637-16 ◽

2016 ◽

Vol 82 (12) ◽

pp. 3671-3682 ◽

Cited By ~ 17

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.

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Interactions between Oral Bacteria: Inhibition of Streptococcus mutans Bacteriocin Production by Streptococcus gordonii

Applied and Environmental Microbiology ◽

10.1128/aem.71.1.354-362.2005 ◽

2005 ◽

Vol 71 (1) ◽

pp. 354-362 ◽

Cited By ~ 90

Author(s):

Bing-Yan Wang ◽

Howard K. Kuramitsu

Keyword(s):

Quorum Sensing ◽

Streptococcus Mutans ◽

Parent Strain ◽

Oral Bacteria ◽

Bacteriocin Production ◽

Oral Streptococci ◽

Streptococcus Gordonii ◽

Signaling System ◽

Signaling Systems ◽

Bacteria Inhibition

ABSTRACT Streptococcus mutans has been recognized as an important etiological agent in human dental caries. Some strains of S. mutans also produce bacteriocins. In this study, we sought to demonstrate that bacteriocin production by S. mutans strains GS5 and BM71 was mediated by quorum sensing, which is dependent on a competence-stimulating peptide (CSP) signaling system encoded by the com genes. We also demonstrated that interactions with some other oral streptococci interfered with S. mutans bacteriocin production both in broth and in biofilms. The inhibition of S. mutans bacteriocin production by oral bacteria was stronger in biofilms than in broth. Using transposon Tn916 mutagenesis, we identified a gene (sgc; named for Streptococcus gordonii challisin) responsible for the inhibition of S. mutans bacteriocin production by S. gordonii Challis. Interruption of the sgc gene in S. gordonii Challis resulted in attenuated inhibition of S. mutans bacteriocin production. The supernatant fluids from the sgc mutant did not inactivate the exogenous S. mutans CSP as did those from the parent strain Challis. S. gordonii Challis did not inactivate bacteriocin produced by S. mutans GS5. Because S. mutans uses quorum sensing to regulate virulence, strategies designed to interfere with these signaling systems may have broad applicability for biological control of this caries-causing organism.

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Comprehensive analysis of bacteriocins in Streptococcus mutans

Scientific Reports ◽

10.1038/s41598-021-92370-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Atsuko Watanabe ◽

Miki Kawada-Matsuo ◽

Mi Nguyen-Tra Le ◽

Junzo Hisatsune ◽

Yuichi Oogai ◽

...

Keyword(s):

Antibacterial Activity ◽

Streptococcus Mutans ◽

Bacterial Species ◽

Comprehensive Analysis ◽

Whole Genome Sequence ◽

Oral Streptococci ◽

Positive Strain ◽

Single Positive ◽

Dental Plaques ◽

Bacteriocin Gene

AbstractStreptococcus mutans produces bacteriocins that show antibacterial activity against several bacteria. However, comprehensive analysis of these bacteriocins has not been well done. In this study, we isolated 125 S. mutans strains from volunteers and determined their whole genome sequence. Based on the genome analysis, the distribution of each bacteriocin gene (mutacins I-IV, K8 and Smb) was investigated. We found 17, 5, and 2 strains showing 100% matches with mutacin I, mutacin II and mutacin III, respectively. Five mutacin III-positive strains had 2 mismatches compared to mature mutacin III. In 67 mutacin IV-positive strains, 38 strains showed 100% match with mutacin IV, while 29 strains showed some variations. In 23 mutacin K8- and 32 mutacin Smb-positive strains, all except one mutacin K8-positive strain showed 100% match with the mature peptides. Among 125 strains, 84 (65.1%), 26 (20.2%), and 5 (3.9%) strains were positive for one, two and three bacteriocin genes, respectively. Then, the antibacterial activity against oral streptococci and other oral bacterial species was investigated by using bacteriocin gene single-positive strains. Each bacteriocin gene-positive strain showed a different pattern of antibacterial activity. These results speculate that individual S. mutans strains may affect the bacterial composition of dental plaques.

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Characterization of Hydrogen Peroxide-Induced DNA Release by Streptococcus sanguinis and Streptococcus gordonii

Journal of Bacteriology ◽

10.1128/jb.00906-09 ◽

2009 ◽

Vol 191 (20) ◽

pp. 6281-6291 ◽

Cited By ~ 70

Author(s):

Jens Kreth ◽

Hung Vu ◽

Yongshu Zhang ◽

Mark C. Herzberg

Keyword(s):

Hydrogen Peroxide ◽

Cell Adhesion ◽

Extracellular Dna ◽

Release Mechanism ◽

Streptococcus Gordonii ◽

Oral Biofilm ◽

Streptococcus Sanguinis ◽

Production Of Hydrogen ◽

Dna Release ◽

Biofilm Development

ABSTRACT Extracellular DNA (eDNA) is produced by several bacterial species and appears to contribute to biofilm development and cell-cell adhesion. We present data showing that the oral commensals Streptococcus sanguinis and Streptococcus gordonii release DNA in a process induced by pyruvate oxidase-dependent production of hydrogen peroxide (H2O2). Surprisingly, S. sanguinis and S. gordonii cell integrity appears unaffected by conditions that cause autolysis in other eDNA-producing bacteria. Exogenous H2O2 causes release of DNA from S. sanguinis and S. gordonii but does not result in obvious lysis of cells. Under DNA-releasing conditions, cell walls appear functionally intact and ribosomes are retained over time. During DNA release, intracellular RNA and ATP are not coreleased. Hence, the release mechanism appears to be highly specific for DNA. Release of DNA without detectable autolysis is suggested to be an adaptation to the competitive oral biofilm environment, where autolysis could create open spaces for competitors to invade. Since eDNA promotes cell-to-cell adhesion, release appears to support oral biofilm formation and facilitates exchange of genetic material among competent strains.

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Eficácia antimicrobiana de ionômeros de vidro restauradores contra espécies bacterianas iniciadoras do biofilme dental e cariogênicas

Revista dos Trabalhos de Iniciação Científica da UNICAMP ◽

10.20396/revpibic2620181458 ◽

2019 ◽

Author(s):

Rafael Nobrega Stipp ◽

Fernanda Felipe de Andrade ◽

Laura da Silva Gabriel ◽

Marina Stênico ◽

Gabriela Castro Fonseca ◽

...

Keyword(s):

Streptococcus Mutans ◽

Streptococcus Gordonii ◽

Streptococcus Sanguinis ◽

Mutans Streptococcus

Os fluoretos liberados dos ionômeros de vidro possuem atividade antimicrobiana, o que contribui para a eficácia do material. O objetivo deste estudo foi avaliar a atividade antimicrobiana de 18 ionômeros convencionais restauradores de vidro contra Streptococcus mutans, Streptococcus gordonii e Streptococcus sanguinis. Foram avaliados a capacidade de inibição do crescimento bacteriano na presença do ionômero e de redução da queda do pH, ocasionada pelos microrganismos estudados. Os iônomeros apresentaram atividade variada em relação aos microrganismos estudados.

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