Journal: Journal of Oral Microbiology

Article Title: Water-insoluble exopolysaccharide synthesized by glucosyltransferases mediates the antibacterial activity of ClyR against Streptococcus mutans

doi: 10.1080/20002297.2025.2566894

Figure Lengend Snippet: The ∆ gtfB mutant lacking GtfB exhibited increased resistance to ClyR. (A) Coomassie-stained SDS-PAGE gel showing the Gtfs content in the culture supernatant of S. mutans UA159 and the ∆ gtfB mutant, with arrows indicating GtfC and GtfD bands. (B) Zymogram assay detecting water-insoluble EPS synthesis activity of Gtfs. Bands corresponding to water-insoluble EPS are labeled. (C) 10-fold serial dilutions of bacteria were spotted onto BHIA plates 1 h and 7 h after treatment with and without ClyR, followed by observation and CFU counting after 48 h. (D) Survival rates of S. mutans UA159 and ∆ gtfB were assessed 1 h and 7 h after exposure to ClyR (* p < 0.05, ** p < 0.01).

Article Snippet: The S. mutans UA159 strain was obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA), and its derivative was provided by the State Key Laboratory of Oral Diseases at Sichuan University.

Techniques: Mutagenesis, Staining, SDS Page, Activity Assay, Labeling, Bacteria

Journal: Journal of Oral Microbiology

Article Title: Water-insoluble exopolysaccharide synthesized by glucosyltransferases mediates the antibacterial activity of ClyR against Streptococcus mutans

doi: 10.1080/20002297.2025.2566894

Figure Lengend Snippet: Biomass and structure of S. mutans UA159 and ∆gtfB biofilms after ClyR treatment. (A) Biofilm morphology and biomass of S. mutans UA159 and ∆ gtfB were examined using SEM (× 20,000 magnification) and CLSM. (B) Quantification of biofilm thickness loss, water-insoluble EPS loss, and bacterial reduction after ClyR treatment for both strains (** p < 0.01).

Article Snippet: The S. mutans UA159 strain was obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA), and its derivative was provided by the State Key Laboratory of Oral Diseases at Sichuan University.

Techniques:

Journal: ACS Omega

Article Title: Putative Prebiotics Can Disrupt 3D Architecture and Modulate the Microbial Population to Prevent Cariogenic Biofilm Build-Up In Vitro

doi: 10.1021/acsomega.5c11573

Figure Lengend Snippet: Biomass of S. mutans biofilms exposed to distinct concentrations of the tested substances and controls. Biomass was inferred from the crystal violet staining assay by measuring the OD at 570 nm. Data are means, and the error bars correspond to 95% confidence interval (CI) ( n = 12). The asterisks (****) correspond to p ≤ 0.0001 (ANOVA one-way, followed by Dunnett’s multiple comparison test). GlcNAc: N -acetyl- d -glucosamine; ARG: arginine; PRO: proline; Control: no substance.

Article Snippet: The strains S. mutans UA159 (ATCC 700610), A. naeslundii ATCC 12104, and S. gordonii DL-1 stocks (at −75 °C in tryptic soy broth plus 20% glycerol) were plated on blood agar plates (37 °C, 10% CO 2 , 48 h).

Techniques: Staining, Comparison, Control

Journal: ACS Omega

Article Title: Putative Prebiotics Can Disrupt 3D Architecture and Modulate the Microbial Population to Prevent Cariogenic Biofilm Build-Up In Vitro

doi: 10.1021/acsomega.5c11573

Figure Lengend Snippet: Effect of tested substances on pH of spent media in S. mutans (A) and mixed-species ( S. mutans , S. gordonii , and A. naeslundii ) (B) biofilms. Data are means, and the error bars correspond to 95% CI. The asterisk (*) represents p ≤ 0.05 for arginine versus all other groups, while the number sign ( # ) represents p ≤ 0.05 for GlcNAc (ANOVA two-way, followed by Tukey’s multiple comparison test). Control: no substance.

Article Snippet: The strains S. mutans UA159 (ATCC 700610), A. naeslundii ATCC 12104, and S. gordonii DL-1 stocks (at −75 °C in tryptic soy broth plus 20% glycerol) were plated on blood agar plates (37 °C, 10% CO 2 , 48 h).

Techniques: Comparison, Control

Journal: ACS Omega

Article Title: Putative Prebiotics Can Disrupt 3D Architecture and Modulate the Microbial Population to Prevent Cariogenic Biofilm Build-Up In Vitro

doi: 10.1021/acsomega.5c11573

Figure Lengend Snippet: Effect of tested substances on S. mutans and mixed-species ( S. mutans , S. gordonii , and A. naeslundii ) biofilms. The figure depicts the viable bacterial population (CFU/biofilm), and the biomass (dry weight: mg/biofilm). The upper graphs represent the data from S. mutans biofilms (A,B), while the bottom graphs represent the data from mixed-species biofilms (C,D). Data are means, and the error bars correspond to 95% CI. The asterisk represents statistical difference for depicted comparisons (**** p ≤ 0.0001; *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05; ANOVA one-way, followed by Tukey’s multiple comparison test for (A–C); ANOVA two-way, followed by Tukey’s multiple comparison test for (D)). Control: no substance.

Article Snippet: The strains S. mutans UA159 (ATCC 700610), A. naeslundii ATCC 12104, and S. gordonii DL-1 stocks (at −75 °C in tryptic soy broth plus 20% glycerol) were plated on blood agar plates (37 °C, 10% CO 2 , 48 h).

Techniques: Comparison, Control

Journal: ACS Omega

Article Title: Putative Prebiotics Can Disrupt 3D Architecture and Modulate the Microbial Population to Prevent Cariogenic Biofilm Build-Up In Vitro

doi: 10.1021/acsomega.5c11573

Figure Lengend Snippet: Microcolonies count (A) and total area occupied (B) in S. mutans single-species and mixed-species biofilms exposed to tested substances. The plotted data represent averages of counts and occupied area of biofilms at 67 h, and error bars indicate 95% CI ( n = 6 per biofilm). The asterisk represents statistical difference for depicted comparisons (**** p ≤ 0.0001; *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05; ANOVA two-way, followed by Tukey’s multiple comparison test). Control: no substance.

Article Snippet: The strains S. mutans UA159 (ATCC 700610), A. naeslundii ATCC 12104, and S. gordonii DL-1 stocks (at −75 °C in tryptic soy broth plus 20% glycerol) were plated on blood agar plates (37 °C, 10% CO 2 , 48 h).

Techniques: Comparison, Control

Journal: ACS Omega

Article Title: Putative Prebiotics Can Disrupt 3D Architecture and Modulate the Microbial Population to Prevent Cariogenic Biofilm Build-Up In Vitro

doi: 10.1021/acsomega.5c11573

Figure Lengend Snippet: Profiles of the distribution of exopolysaccharides and bacteria in S. mutans single-species biofilms exposed to tested substances. The data represent the mean percentage of coverage per area, ranging from the interface of the HA disc-biofilm to the outer layer of each biofilm at 67 h (error bars indicate 95% CI; n = 6 images per biofilm). The vertical line serves as a guide to compare the tested substances with the control, which contains no substance. Control: no substance.

Article Snippet: The strains S. mutans UA159 (ATCC 700610), A. naeslundii ATCC 12104, and S. gordonii DL-1 stocks (at −75 °C in tryptic soy broth plus 20% glycerol) were plated on blood agar plates (37 °C, 10% CO 2 , 48 h).

Techniques: Bacteria, Control

Journal: ACS Omega

Article Title: Putative Prebiotics Can Disrupt 3D Architecture and Modulate the Microbial Population to Prevent Cariogenic Biofilm Build-Up In Vitro

doi: 10.1021/acsomega.5c11573

Figure Lengend Snippet: Architecture of S. mutans biofilms exposed to tested substances. Representative 3-D renderings of biofilms at 67 h of development. Green represents bacterial cells labeled with SYTO 9, and red represents the exopolysaccharides stained with Alexa Fluor 647-labeled dextran conjugate. The imaging was performed using a Zeiss LSM 780 AxioObserver microscope equipped with a 20×/0.5 objective lens. The larger image in each set represents the overlaid red and green channels. The scale bar corresponds to 50 μm. Control: no substance.

Article Snippet: The strains S. mutans UA159 (ATCC 700610), A. naeslundii ATCC 12104, and S. gordonii DL-1 stocks (at −75 °C in tryptic soy broth plus 20% glycerol) were plated on blood agar plates (37 °C, 10% CO 2 , 48 h).

Techniques: Labeling, Staining, Imaging, Microscopy, Control

Journal: ACS Omega

Article Title: Putative Prebiotics Can Disrupt 3D Architecture and Modulate the Microbial Population to Prevent Cariogenic Biofilm Build-Up In Vitro

doi: 10.1021/acsomega.5c11573

Figure Lengend Snippet: Biovolume of exopolysaccharides and bacteria in S. mutans single-species and mixed-species biofilms exposed to the tested substances. Biovolumes are represented as the biomasses (μm 3 /μm 2 ) of bacteria and exopolysaccharides for S. mutans (A) and mixed-species (B) biofilms at 67 h. These data were used to determine the ratio of bacterial biomass (biovolume) to exopolysaccharides (C). The plotted data represent averages, and error bars indicate 95% CI ( n = 6 per biofilm). The asterisk represents statistical difference for depicted comparisons (**** p ≤ 0.0001; *** p ≤ 0.001; ** p ≤ 0.01; ANOVA two-way, followed by Tukey’s multiple comparison test). Control: no substance.

Article Snippet: The strains S. mutans UA159 (ATCC 700610), A. naeslundii ATCC 12104, and S. gordonii DL-1 stocks (at −75 °C in tryptic soy broth plus 20% glycerol) were plated on blood agar plates (37 °C, 10% CO 2 , 48 h).

Techniques: Bacteria, Comparison, Control

Journal: Microbiology Spectrum

Article Title: Inhibitory effects of berberine against Streptococcus mutans : an in vitro insight on its anticaries potential

doi: 10.1128/spectrum.00700-25

Figure Lengend Snippet: Inhibitory effect of BBR on the growth of bacteria. ( A ) The MIC and MBC values of BBR against planktonic bacteria. 10 6 CFU/mL bacteria in BHI broth were incubated with final concentrations of BBR ranging from 25 to 400 μg/mL anaerobically at 37°C for 24 h. The MIC was defined as the lowest concentration of BBR that inhibited visible bacterial growth. At the termination of the MIC assay, a volume of the culture was struck on BHIA and incubated to observe growth. The MBC was defined as the lowest concentration that yielded no colony growth by subculturing on BHIA plates. ( B–F ) The 24 h growth curve of planktonic S. mutans UA159, S. mutans ATCC 25175, S. mutans GS-5, S. gordonii DL-1, and S. mitis ATCC 6249 incubated under anaerobic conditions with/without treatment of BBR in a 96-well plate. The absorbance of each well was recorded every hour. ( G and H ) The short-term antibacterial effect of BBR on S. mutans UA159 was assessed by measuring the average number of CFU. S. mutans UA159 was treated with BBR for 5 min ( G ) and 10 min ( H ). The suspensions were then diluted and plated onto BHIA plates and incubated under anaerobic conditions at 37°C for 24 h to determine CFU counts. Values represent the means ± SD from three independent experiments (** P ＜0.01, *** P ＜0.001).

Article Snippet: The bacterial strain S. mutans UA159 was obtained from the American Type Culture Collection (ATCC).

Techniques: Bacteria, Incubation, Concentration Assay, Subculturing Assay

Journal: Microbiology Spectrum

Article Title: Inhibitory effects of berberine against Streptococcus mutans : an in vitro insight on its anticaries potential

doi: 10.1128/spectrum.00700-25

Figure Lengend Snippet: Effect of BBR on the biofilm formation of S. mutans . In these assays, BBR was incubated with S. mutans in BHIS for 24 h to evaluate its effect on the biofilm formation. ( A ) The crystal violet staining of S. mutans 24 h biofilms under the treatment of BBR. The images were taken from a 24-well cell culture plate. ( B ) The results of the crystal violet staining assay applied to S. mutans biofilms after treated with different concentrations of BBR for 24 h in BHIS. ( C ) SEM micro-images of S. mutans 24 h biofilms on glass coverslips. In the control group, S. mutans bacteria formed dense biofilms covered with large amounts of EPS (red arrows). Disrupted biofilm skeleton (yellow arrows) and enlarged biofilm pores (green arrows) were observed under the treatment of 50 µg/mL of BBR. Only a small number of morphologically irregular S. mutans and cell contents were observed under the treatment of 100 µg/mL and 200 µg/mL of BBR (purple arrows). Values represent the means ± SD from three independent experiments (*** P ＜0.001).

Article Snippet: The bacterial strain S. mutans UA159 was obtained from the American Type Culture Collection (ATCC).

Techniques: Incubation, Staining, Cell Culture, Control, Bacteria

Journal: Microbiology Spectrum

Article Title: Inhibitory effects of berberine against Streptococcus mutans : an in vitro insight on its anticaries potential

doi: 10.1128/spectrum.00700-25

Figure Lengend Snippet: Quantification of S. mutans biofilm after treatment of BBR. ( A ) Double-labeling imaging of S. mutans 24 h biofilms formed on glass coverslips. Live bacteria were green-labeled, and dead bacteria were red-labeled. ( B ) BBR reduced the proportion of live bacteria in S. mutans biofilms. ( C ) The effect of BBR on S. mutans biofilm formation was assessed by measuring the average number of CFU in the biofilm in one well of the culture plate. The analysis was performed using Image J COMSTAT software. Values represent the means ± SD from three independent experiments (*** P ＜0.001).

Article Snippet: The bacterial strain S. mutans UA159 was obtained from the American Type Culture Collection (ATCC).

Techniques: Labeling, Imaging, Bacteria, Software

Journal: Microbiology Spectrum

Article Title: Inhibitory effects of berberine against Streptococcus mutans : an in vitro insight on its anticaries potential

doi: 10.1128/spectrum.00700-25

Figure Lengend Snippet: Effect of BBR on eradicating the S. mutans mature biofilms. ( A ) Double-labeling imaging of S. mutans mature biofilms treated with BBR. Live and dead bacteria were green-labeled and red-labeled, respectively. ( B ) BBR decreases the proportion of live bacteria in S. mutans mature biofilms. ( C ) The effect of BBR on eradicating the S. mutans mature biofilms was measured by the determination of the average number of CFUs in the biofilm. The S. mutans biofilms were cultured for 24 h without BBR. Then, BBR solution was added to each well for another 24 h. The data were collected at 48 h. The analysis was performed using Image J COMSTAT software. Values represent the means ± SD from three independent experiments (*** P ＜0.001).

Article Snippet: The bacterial strain S. mutans UA159 was obtained from the American Type Culture Collection (ATCC).

Techniques: Labeling, Imaging, Bacteria, Cell Culture, Software

Journal: Microbiology Spectrum

Article Title: Inhibitory effects of berberine against Streptococcus mutans : an in vitro insight on its anticaries potential

doi: 10.1128/spectrum.00700-25

Figure Lengend Snippet: BBR inhibited S. mutans virulence factors. ( A ) Effect of BBR on the biofilm structure of S. mutans observed by CLSM. Double-labeling imaging of S. mutans 24 h biofilm formed on glass coverslips. The fluorescence (SYTO 9) marks the live bacteria, while the red fluorescence (Concanavalin A-TRITC) marks the EPS synthesized by S. mutans . ( B ) Quantification of the amounts of EPS and bacteria in each scanned layer of S. mutans 24 h biofilm without the treatment of BBR. ( C ) Quantification of the amounts of EPS and bacteria in each scanned layer of S. mutans 24 h biofilm with the treatment of 50 µg/mL BBR. ( D ) Quantitative measurement of water-insoluble EPS by anthrone-sulfuric method. ( E ) Measurement of lactic acid production. ( F and G ) Effect of BBR on S. mutans glycolytic pH drop under 1% ( F ) and 0.1% ( G ) glucose. Values represent the means ± SD from three independent experiments (*** P ＜0.001).

Article Snippet: The bacterial strain S. mutans UA159 was obtained from the American Type Culture Collection (ATCC).

Techniques: Labeling, Imaging, Fluorescence, Bacteria, Synthesized

Journal: Microbiology Spectrum

Article Title: Inhibitory effects of berberine against Streptococcus mutans : an in vitro insight on its anticaries potential

doi: 10.1128/spectrum.00700-25

Figure Lengend Snippet: The bactericidal mechanism of BBR on S. mutans . ( A ) Effect of BBR on aggregation. ( B ) Membrane depolarization activity of BBR was tested using DiSC3(5). ( C ) TEM micrographs of S. mutans . Values represent the means ± SD from three independent experiments (* P ＜0.05, *** P ＜0.001).

Article Snippet: The bacterial strain S. mutans UA159 was obtained from the American Type Culture Collection (ATCC).

Techniques: Membrane, Activity Assay

Journal: Microbiology Spectrum

Article Title: Inhibitory effects of berberine against Streptococcus mutans : an in vitro insight on its anticaries potential

doi: 10.1128/spectrum.00700-25

Figure Lengend Snippet: Effects of BBR on S. mutans gene expression. The relative mRNA expressions of gtfB , gtfC , gtfD , ldh , vicR , liaR , and comD were measured by qRT-PCR. S. mutans UA159 16S rRNA was used as an internal control. Values represent the means ± SD from three independent experiments (*** P ＜0.001, ns, not statistically significant compared to the untreated control group).

Article Snippet: The bacterial strain S. mutans UA159 was obtained from the American Type Culture Collection (ATCC).

Techniques: Gene Expression, Quantitative RT-PCR, Control

Journal: Microbiology Spectrum

Article Title: Inhibitory effects of berberine against Streptococcus mutans : an in vitro insight on its anticaries potential

doi: 10.1128/spectrum.00700-25

Figure Lengend Snippet: Sequences of primers used in this study

Article Snippet: The bacterial strain S. mutans UA159 was obtained from the American Type Culture Collection (ATCC).

Techniques: