Structured Review

Fisher Scientific biofilms
Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. <t>Biofilms</t> were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p
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1) Product Images from "Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions"

Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.00258

Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. Biofilms were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p
Figure Legend Snippet: Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. Biofilms were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p

Techniques Used: Sonication, Polymerase Chain Reaction

Candida albicans mycofilms facilitates Staphylococcus aureus biofilm formation. Mono- and dual-species biofilms were standardized 1 × 10 6 CFU/mL and grown on 96-well plates for 90 min, 6 and 24 h. Biofilms were then washed with phosphate buffered saline (PBS) and biomass assessed using the crystal violet assay (A). Standardized biofilms were grown before being sonicated to remove the biomass. Live/dead PCR was then used to extract DNA and determine total and live colony forming equivalents (CFE) from mono- and dual-species biofilms (B) . Biofilm morphology was then analyzed using CSLM. Biofilms were grown before being fluorescently stained using calcofluor white and SYTO9 ® dyes. Resulting biofilms were then viewed on a Leica SP5 laser scanning confocal microscope and images were then processed and analyzed using Volocity 3D Image Analysis Software (C) . Results represent data from three independent occasions. Statistical analysis compares dual-species biofilms to their mono-species equivalent ( ∗ p
Figure Legend Snippet: Candida albicans mycofilms facilitates Staphylococcus aureus biofilm formation. Mono- and dual-species biofilms were standardized 1 × 10 6 CFU/mL and grown on 96-well plates for 90 min, 6 and 24 h. Biofilms were then washed with phosphate buffered saline (PBS) and biomass assessed using the crystal violet assay (A). Standardized biofilms were grown before being sonicated to remove the biomass. Live/dead PCR was then used to extract DNA and determine total and live colony forming equivalents (CFE) from mono- and dual-species biofilms (B) . Biofilm morphology was then analyzed using CSLM. Biofilms were grown before being fluorescently stained using calcofluor white and SYTO9 ® dyes. Resulting biofilms were then viewed on a Leica SP5 laser scanning confocal microscope and images were then processed and analyzed using Volocity 3D Image Analysis Software (C) . Results represent data from three independent occasions. Statistical analysis compares dual-species biofilms to their mono-species equivalent ( ∗ p

Techniques Used: Crystal Violet Assay, Sonication, Polymerase Chain Reaction, Staining, Microscopy, Software

Scanning electron micrograph of S. aureus colonizing C. albicans hyphae within dual-species biofilms. Dual-species biofilms were grown for 24 h before being fixed, processed, and imaged using a JEOL-JSM 6400 scanning electron microscope. S. aureus colonies can be seen adhering and embedded within the hyphal meshwork of C. albicans . White arrows indicate clusters of S. aureus colonies encased within extracellular matrix (ECM). Scale bar represents 5 μm at × 5000 magnification.
Figure Legend Snippet: Scanning electron micrograph of S. aureus colonizing C. albicans hyphae within dual-species biofilms. Dual-species biofilms were grown for 24 h before being fixed, processed, and imaged using a JEOL-JSM 6400 scanning electron microscope. S. aureus colonies can be seen adhering and embedded within the hyphal meshwork of C. albicans . White arrows indicate clusters of S. aureus colonies encased within extracellular matrix (ECM). Scale bar represents 5 μm at × 5000 magnification.

Techniques Used: Microscopy

Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p
Figure Legend Snippet: Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p

Techniques Used: SYBR Green Assay, Fluorescence, Real-time Polymerase Chain Reaction, Staining, Incubation

2) Product Images from "Nontypeable Haemophilus influenzae Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate In Vivo, Which Can Be Used as a Target for Bactericidal Antibody"

Article Title: Nontypeable Haemophilus influenzae Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate In Vivo, Which Can Be Used as a Target for Bactericidal Antibody

Journal: mBio

doi: 10.1128/mBio.01401-18

Confocal micrograph of a biofilm formed in a chinchilla middle ear by NTHi strain 2019 stained with MAb 6E4. The antibody was counterstained with a goat anti-mouse anti IgG antibody conjugated to fluorescein. The DNA in the biofilm was counterstained with DAPI and fluoresces blue. Organisms can be seen throughout the biofilm stained with MAb 6E4, indicating the presence of the KDO epitope in vivo .
Figure Legend Snippet: Confocal micrograph of a biofilm formed in a chinchilla middle ear by NTHi strain 2019 stained with MAb 6E4. The antibody was counterstained with a goat anti-mouse anti IgG antibody conjugated to fluorescein. The DNA in the biofilm was counterstained with DAPI and fluoresces blue. Organisms can be seen throughout the biofilm stained with MAb 6E4, indicating the presence of the KDO epitope in vivo .

Techniques Used: Staining, In Vivo

3) Product Images from "Rhesus θ-defensin-1 (RTD-1) exhibits in vitro and in vivo activity against cystic fibrosis strains of Pseudomonas aeruginosa"

Article Title: Rhesus θ-defensin-1 (RTD-1) exhibits in vitro and in vivo activity against cystic fibrosis strains of Pseudomonas aeruginosa

Journal: Journal of Antimicrobial Chemotherapy

doi: 10.1093/jac/dkv301

Activity of RTD-1 against PAO1 biofilms. Biofilms were grown on polycarbonate membranes placed on M63 agarose plates as described in the Materials and methods section. (a) Electron microscopy image of PAO1 aggregates on polycarbonate membranes. After
Figure Legend Snippet: Activity of RTD-1 against PAO1 biofilms. Biofilms were grown on polycarbonate membranes placed on M63 agarose plates as described in the Materials and methods section. (a) Electron microscopy image of PAO1 aggregates on polycarbonate membranes. After

Techniques Used: Activity Assay, Electron Microscopy

4) Product Images from "Candida albicans Biofilms Produce Antifungal-Tolerant Persister Cells ▿"

Article Title: Candida albicans Biofilms Produce Antifungal-Tolerant Persister Cells ▿

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00684-06

Isolation of persister cells from a biofilm. C. albicans MC191 was grown as a biofilm for 48 h in RPMI 1640 medium in microtiter plate wells. A homogenous population of cells from disrupted biofilms was obtained by applying a forward scatter and a side scatter gate, as shown in panel A, for all subsequent analyses. (B) A biofilm was stained with 100 μg/ml fluorescein diacetate for 24 h, disrupted, washed three times with PBS, and analyzed with a MoFlo cell sorter. Single events representing individual cells were physically sorted directly on YPD medium and incubated for 48 h (D). (C) A biofilm was treated with 100 μg/ml amphotericin B, stained with fluorescein diacetate, and similarly analyzed with the cell sorter. Two distinct populations were separated based on green fluorescence intensity, as shown. (E) Particles representing 96 events from the dim population, R4, were sorted onto YPD agar and incubated for 48 h. (F) Particles representing over 6,000 events from R2 were sorted onto YPD agar and incubated for 48 h.
Figure Legend Snippet: Isolation of persister cells from a biofilm. C. albicans MC191 was grown as a biofilm for 48 h in RPMI 1640 medium in microtiter plate wells. A homogenous population of cells from disrupted biofilms was obtained by applying a forward scatter and a side scatter gate, as shown in panel A, for all subsequent analyses. (B) A biofilm was stained with 100 μg/ml fluorescein diacetate for 24 h, disrupted, washed three times with PBS, and analyzed with a MoFlo cell sorter. Single events representing individual cells were physically sorted directly on YPD medium and incubated for 48 h (D). (C) A biofilm was treated with 100 μg/ml amphotericin B, stained with fluorescein diacetate, and similarly analyzed with the cell sorter. Two distinct populations were separated based on green fluorescence intensity, as shown. (E) Particles representing 96 events from the dim population, R4, were sorted onto YPD agar and incubated for 48 h. (F) Particles representing over 6,000 events from R2 were sorted onto YPD agar and incubated for 48 h.

Techniques Used: Isolation, Staining, Incubation, Fluorescence

Survival of C. albicans biofilms challenged with amphotericin B and chlorhexidine. Biofilms were treated with 100 μg/ml amphotericin B, 100 μg/ml chlorhexidine, or a combination of the two antifungals for 24 h. The biofilms were washed and sampled for CFU determination before and after antibiotic treatment. The experiment was performed in triplicate, and the error bars indicate standard deviations.
Figure Legend Snippet: Survival of C. albicans biofilms challenged with amphotericin B and chlorhexidine. Biofilms were treated with 100 μg/ml amphotericin B, 100 μg/ml chlorhexidine, or a combination of the two antifungals for 24 h. The biofilms were washed and sampled for CFU determination before and after antibiotic treatment. The experiment was performed in triplicate, and the error bars indicate standard deviations.

Techniques Used:

Survival of C. albicans 3153A biofilm and exponential- and stationary-phase cells. Biofilms were cultured in RPMI medium for 48 h, scraped, vortexed, resuspended in 100 μl PBS, and plated for CFU determination. Exponential- and stationary-phase cultures were obtained by growth in the same medium. The experiment was performed in triplicate, and the error bars represent standard deviations. (A) Amphotericin B. (B) Chlorhexidine.
Figure Legend Snippet: Survival of C. albicans 3153A biofilm and exponential- and stationary-phase cells. Biofilms were cultured in RPMI medium for 48 h, scraped, vortexed, resuspended in 100 μl PBS, and plated for CFU determination. Exponential- and stationary-phase cultures were obtained by growth in the same medium. The experiment was performed in triplicate, and the error bars represent standard deviations. (A) Amphotericin B. (B) Chlorhexidine.

Techniques Used: Cell Culture

Heritability of persister formation. Biofilms were treated with 100 μg/ml amphotericin B or 100 μg/ml chlorhexidine for 24 h, after which they were disrupted by being vortexed, washed, and reinoculated in order to form new biofilms. The biofilms were sampled for CFU determination before and after antibiotic treatment. The procedure was repeated a total of three times. The experiment was performed in triplicate, and the error bars indicate standard deviations.
Figure Legend Snippet: Heritability of persister formation. Biofilms were treated with 100 μg/ml amphotericin B or 100 μg/ml chlorhexidine for 24 h, after which they were disrupted by being vortexed, washed, and reinoculated in order to form new biofilms. The biofilms were sampled for CFU determination before and after antibiotic treatment. The procedure was repeated a total of three times. The experiment was performed in triplicate, and the error bars indicate standard deviations.

Techniques Used:

Live-dead staining of C. albicans with fluorescein diacetate. Planktonic or biofilm cells were stained with 100 μg/ml fluorescein diacetate and examined by fluorescence microscopy. (A) Live planktonic cells. (B) Dead planktonic cells after treatment with 100 μg/ml amphotericin B (×400 magnification). (C, D, and E) Biofilms (×1,000 magnification) of untreated control and after 18 and 48 h of amphotericin B treatment (100 μg/ml), respectively.
Figure Legend Snippet: Live-dead staining of C. albicans with fluorescein diacetate. Planktonic or biofilm cells were stained with 100 μg/ml fluorescein diacetate and examined by fluorescence microscopy. (A) Live planktonic cells. (B) Dead planktonic cells after treatment with 100 μg/ml amphotericin B (×400 magnification). (C, D, and E) Biofilms (×1,000 magnification) of untreated control and after 18 and 48 h of amphotericin B treatment (100 μg/ml), respectively.

Techniques Used: Staining, Fluorescence, Microscopy

5) Product Images from "Genome-Wide Transcription Profiling of the Early Phase of Biofilm Formation by Candida albicans †"

Article Title: Genome-Wide Transcription Profiling of the Early Phase of Biofilm Formation by Candida albicans †

Journal: Eukaryotic Cell

doi: 10.1128/EC.4.9.1562-1573.2005

Phloxine B staining of C. albicans ) and photographed (Arcturus). Differences in staining are evident during the 6 h of biofilm development. (A) T = 30; (B) T = 90; (C) T = 270. Magnification, ×40.
Figure Legend Snippet: Phloxine B staining of C. albicans ) and photographed (Arcturus). Differences in staining are evident during the 6 h of biofilm development. (A) T = 30; (B) T = 90; (C) T = 270. Magnification, ×40.

Techniques Used: Staining

). The scan rates were about 1 Hz with scan sizes between 15 and 60 μm. (A) Cell adhesion ( T = 30); (B to E) biofilm development ( T = 60 to T = 390). Bar: 10 μm (A and B) and 30 μm (C to E).
Figure Legend Snippet: ). The scan rates were about 1 Hz with scan sizes between 15 and 60 μm. (A) Cell adhesion ( T = 30); (B to E) biofilm development ( T = 60 to T = 390). Bar: 10 μm (A and B) and 30 μm (C to E).

Techniques Used:

6) Product Images from "The Cyclic AMP-Dependent Catabolite Repression System of Serratia marcescens Mediates Biofilm Formation through Regulation of Type 1 Fimbriae "

Article Title: The Cyclic AMP-Dependent Catabolite Repression System of Serratia marcescens Mediates Biofilm Formation through Regulation of Type 1 Fimbriae

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.02733-07

S. marcescens catabolite repression genes regulate biofilm formation and cAMP production. (A) Biofilms formed on test tubes that were grown at 30°C on a rotor for 14 to 15 h. Results are shown for mutation and complementation of cyaA , crr , and
Figure Legend Snippet: S. marcescens catabolite repression genes regulate biofilm formation and cAMP production. (A) Biofilms formed on test tubes that were grown at 30°C on a rotor for 14 to 15 h. Results are shown for mutation and complementation of cyaA , crr , and

Techniques Used: Mutagenesis

Glucose stimulates S. marcescens biofilm formation. (A) Photographs of crystal violet-stained biofilms (black arrow) formed on glass test tubes. Biofilms were formed in at 30°C for 15 h under high-sheer conditions in LB supplemented with glucose
Figure Legend Snippet: Glucose stimulates S. marcescens biofilm formation. (A) Photographs of crystal violet-stained biofilms (black arrow) formed on glass test tubes. Biofilms were formed in at 30°C for 15 h under high-sheer conditions in LB supplemented with glucose

Techniques Used: Staining

Catabolite repression proteins regulate S. marcescens biofilm formation.
Figure Legend Snippet: Catabolite repression proteins regulate S. marcescens biofilm formation.

Techniques Used:

Fimbriae are necessary for catabolite repression mutant biofilm phenotypes. (A) Mutation of fimC suppressed the hyperbiofilm phenotype caused by high levels of glucose or mutation of catabolite repression genes. For panels B and C, a kinetic assessment
Figure Legend Snippet: Fimbriae are necessary for catabolite repression mutant biofilm phenotypes. (A) Mutation of fimC suppressed the hyperbiofilm phenotype caused by high levels of glucose or mutation of catabolite repression genes. For panels B and C, a kinetic assessment

Techniques Used: Mutagenesis

7) Product Images from "Role of de-N-acetylase PgaB from Aggregatibacter actinomycetemcomitans in exopolysaccharide export in biofilm mode of growth"

Article Title: Role of de-N-acetylase PgaB from Aggregatibacter actinomycetemcomitans in exopolysaccharide export in biofilm mode of growth

Journal: Molecular oral microbiology

doi: 10.1111/omi.12188

The role of the catalytic domain of PgaB in phenotypic variation and biofilm integrity. (a) Deletion of catalytic domain of PgaB (ΔNpgaB) results in a phenotype with attenuated rugged edges characteristic of IDH 781 but lacking the internal star
Figure Legend Snippet: The role of the catalytic domain of PgaB in phenotypic variation and biofilm integrity. (a) Deletion of catalytic domain of PgaB (ΔNpgaB) results in a phenotype with attenuated rugged edges characteristic of IDH 781 but lacking the internal star

Techniques Used:

Confocal scanning laser microscopic image of biofilms. (a) Biofilm growth was imaged at seven different locations and averaged for analysis. The scale bar is at 20 μm; (b) Cell viability for IDH 781 and ΔNpgaB strains at different time
Figure Legend Snippet: Confocal scanning laser microscopic image of biofilms. (a) Biofilm growth was imaged at seven different locations and averaged for analysis. The scale bar is at 20 μm; (b) Cell viability for IDH 781 and ΔNpgaB strains at different time

Techniques Used:

Immunofluorescence evaluation of biofilms for the production of PNAG. PNAG was visualized using the human mAb F598 and a secondary anti-human IgG conjugated to Alexa 488 (left panel). Images of the same field viewed by DAPI to stain DNA is shown in the
Figure Legend Snippet: Immunofluorescence evaluation of biofilms for the production of PNAG. PNAG was visualized using the human mAb F598 and a secondary anti-human IgG conjugated to Alexa 488 (left panel). Images of the same field viewed by DAPI to stain DNA is shown in the

Techniques Used: Immunofluorescence, Staining

8) Product Images from "In Vitro Analyses of the Effects of Heparin and Parabens on Candida albicans Biofilms and Planktonic Cells"

Article Title: In Vitro Analyses of the Effects of Heparin and Parabens on Candida albicans Biofilms and Planktonic Cells

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.05061-11

In vitro effects of heparin sodium preparation and individual reagents against mature biofilms of and formation of biofilms by C. albicans reference strain SC5314. (A) Effect of each reagent tested on mature biofilms formed by the reference clinical strain
Figure Legend Snippet: In vitro effects of heparin sodium preparation and individual reagents against mature biofilms of and formation of biofilms by C. albicans reference strain SC5314. (A) Effect of each reagent tested on mature biofilms formed by the reference clinical strain

Techniques Used: In Vitro

In vitro effects of the heparin sodium preparation and individual reagents against mature C. albicans biofilms. Mature biofilms were incubated with RPMI 1640 containing serial dilutions (1× to 10×) of each reagent as indicated. The mean
Figure Legend Snippet: In vitro effects of the heparin sodium preparation and individual reagents against mature C. albicans biofilms. Mature biofilms were incubated with RPMI 1640 containing serial dilutions (1× to 10×) of each reagent as indicated. The mean

Techniques Used: In Vitro, Incubation

In vitro effects of heparin sodium preparation and individual reagents against C. albicans biofilm formation. Planktonic cells were resuspended at a density of 1 × 10 6 cells ml −1 in RPMI 1640 medium containing serial dilutions (1×
Figure Legend Snippet: In vitro effects of heparin sodium preparation and individual reagents against C. albicans biofilm formation. Planktonic cells were resuspended at a density of 1 × 10 6 cells ml −1 in RPMI 1640 medium containing serial dilutions (1×

Techniques Used: In Vitro

Structural effects of heparin and parabens on biofilm formation. Sample preparation for scanning electron microscopy was performed on biofilm samples formed on a coverslip after 24 h of incubation of a 0.5-ml inoculum containing 1 × 10 6 cells
Figure Legend Snippet: Structural effects of heparin and parabens on biofilm formation. Sample preparation for scanning electron microscopy was performed on biofilm samples formed on a coverslip after 24 h of incubation of a 0.5-ml inoculum containing 1 × 10 6 cells

Techniques Used: Sample Prep, Electron Microscopy, Incubation

9) Product Images from "A Novel Regulator Modulates Glucan Production, Cell Aggregation and Biofilm Formation in Streptococcus sanguinis SK36"

Article Title: A Novel Regulator Modulates Glucan Production, Cell Aggregation and Biofilm Formation in Streptococcus sanguinis SK36

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.01154

The fiber-like matrix in the biofilms of WT and Δ brpL . The 1-day biofilms of WT (A) and Δ brpL (B) were captured by SEM with magnifications of 100×, 1000×, and 10000×, respectively. Scale bars were shown on each image. The red arrow indicates fiber-like matrix at the periphery of cell aggregation and the blue arrow points to cells inside of the aggregate.
Figure Legend Snippet: The fiber-like matrix in the biofilms of WT and Δ brpL . The 1-day biofilms of WT (A) and Δ brpL (B) were captured by SEM with magnifications of 100×, 1000×, and 10000×, respectively. Scale bars were shown on each image. The red arrow indicates fiber-like matrix at the periphery of cell aggregation and the blue arrow points to cells inside of the aggregate.

Techniques Used:

The effect of SSA_0222 on growth and biofilm formation. (A) The expression of SSA_0222 in WT and Δ brpL was tested by qRT-PCR. The relative expression of SSA_0222 was shown. (B) Strains were cultured in BM supplemented with different kinds of carbohydrate for 24 h. Cell growth was monitored at 600 nm with a plate reader. (C) The 1-day biofilms of strains were quantified by CV staining. All of the data were relative to their WT controls. ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown.
Figure Legend Snippet: The effect of SSA_0222 on growth and biofilm formation. (A) The expression of SSA_0222 in WT and Δ brpL was tested by qRT-PCR. The relative expression of SSA_0222 was shown. (B) Strains were cultured in BM supplemented with different kinds of carbohydrate for 24 h. Cell growth was monitored at 600 nm with a plate reader. (C) The 1-day biofilms of strains were quantified by CV staining. All of the data were relative to their WT controls. ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown.

Techniques Used: Expressing, Quantitative RT-PCR, Cell Culture, Staining

The characteristic of biomass and cell distribution in the biofilms of WT and Δ brpL. (A) The biofilms of WT and Δ brpL were cultured in a 4-well chamber for 24 h, stained by SYTO 9 (green)/PI (red) and captured by CLSM. 3D architectures of biofilms were shown on the top. Biomass (the signal of SYTO 9) and PI signal representing dead cells and eDNA in CLSM images were calculated by COMSTAT analysis (mean ± SD). Average and maximum thickness were quantified by COMSTAT based on the signal of SYTO 9 (mean ± SD). (B) Heat maps showing the thickness of biofilms (overlap of biomass in all slices) were made by COMSTAT analysis, which reflected the distribution of biomass in biofilms. Blue arrows point to cell aggregation in the biofilm of Δ brpL . Scale bars were indicated on the corresponding images. All the data in (A) are compared with their WT control. ∗∗ P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown.
Figure Legend Snippet: The characteristic of biomass and cell distribution in the biofilms of WT and Δ brpL. (A) The biofilms of WT and Δ brpL were cultured in a 4-well chamber for 24 h, stained by SYTO 9 (green)/PI (red) and captured by CLSM. 3D architectures of biofilms were shown on the top. Biomass (the signal of SYTO 9) and PI signal representing dead cells and eDNA in CLSM images were calculated by COMSTAT analysis (mean ± SD). Average and maximum thickness were quantified by COMSTAT based on the signal of SYTO 9 (mean ± SD). (B) Heat maps showing the thickness of biofilms (overlap of biomass in all slices) were made by COMSTAT analysis, which reflected the distribution of biomass in biofilms. Blue arrows point to cell aggregation in the biofilm of Δ brpL . Scale bars were indicated on the corresponding images. All the data in (A) are compared with their WT control. ∗∗ P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown.

Techniques Used: Cell Culture, Staining, Confocal Laser Scanning Microscopy

The gtfP gene is important for Δ brpL to regulate biofilm formation. (A) The biofilms were stained by SYTO 9 (green)/PI (red), visualized by CLSM and quantified by COMSTAT. Biofilm biomass was shown. (B) The heat map of biomass distribution in the biofilm of Δ brpL Δ gtfP was made by COMSTAT analysis. A scale bar was shown on the image. (C) qRT-PCR was performed to examine the expression of gtfP gene in Δ brpL . (D) The WIG and WSG of WT and Δ brpL were measured by the method described in section “Materials and Methods.” ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown. ND indicates no significant difference.
Figure Legend Snippet: The gtfP gene is important for Δ brpL to regulate biofilm formation. (A) The biofilms were stained by SYTO 9 (green)/PI (red), visualized by CLSM and quantified by COMSTAT. Biofilm biomass was shown. (B) The heat map of biomass distribution in the biofilm of Δ brpL Δ gtfP was made by COMSTAT analysis. A scale bar was shown on the image. (C) qRT-PCR was performed to examine the expression of gtfP gene in Δ brpL . (D) The WIG and WSG of WT and Δ brpL were measured by the method described in section “Materials and Methods.” ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown. ND indicates no significant difference.

Techniques Used: Staining, Confocal Laser Scanning Microscopy, Quantitative RT-PCR, Expressing

The biofilm attachment of WT and Δ brpL on polystyrene microtiter plates. The 1-day biofilms of WT and Δ brpL were quantified by CV staining. The washing steps were done by using a Caliper Sciclone G3 liquid handling robot with different washing speeds. P -values were generated by Student’s t- test. Means and standard deviations from triplicate experiments are shown.
Figure Legend Snippet: The biofilm attachment of WT and Δ brpL on polystyrene microtiter plates. The 1-day biofilms of WT and Δ brpL were quantified by CV staining. The washing steps were done by using a Caliper Sciclone G3 liquid handling robot with different washing speeds. P -values were generated by Student’s t- test. Means and standard deviations from triplicate experiments are shown.

Techniques Used: Staining, Generated

The mechanism by which BrpL modulates biofilm formation in Streptococcus sanguinis SK36. GtfP, a glucosyltransferase, is responsible for glucan production. SSA_0222, a component of a PTS, may play an important role in the uptake of sucrose and is essential for survival in BM supplemented with 1% sucrose. BrpL regulates the expression of gtfP and SSA_0222 through the inhibition of another biofilm-related regulator ciaR and as a result controls the biofilm formation of S. sanguinis SK36.
Figure Legend Snippet: The mechanism by which BrpL modulates biofilm formation in Streptococcus sanguinis SK36. GtfP, a glucosyltransferase, is responsible for glucan production. SSA_0222, a component of a PTS, may play an important role in the uptake of sucrose and is essential for survival in BM supplemented with 1% sucrose. BrpL regulates the expression of gtfP and SSA_0222 through the inhibition of another biofilm-related regulator ciaR and as a result controls the biofilm formation of S. sanguinis SK36.

Techniques Used: Expressing, Inhibition

BrpL controls biofilm formation via a downstream regulator CiaR. (A) qRT-PCR was performed to examine the relationships of regulation between ciaR, brpL , and brpT . (B) ), the number of overlapped genes with differential expression (fold change ≥ 1.5 or ≤ 0.667 and p -value ≤ 0.05) in Δ brpL and Δ ciaR was shown by Venn diagram. (C) The expression of genes in WT, Δ brpL and Δ brpL Δ ciaR was tested by qRT-PCR. All of the data were relative to their WT controls. (D) The biofilm biomass of strains was measured by CV staining. ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown.
Figure Legend Snippet: BrpL controls biofilm formation via a downstream regulator CiaR. (A) qRT-PCR was performed to examine the relationships of regulation between ciaR, brpL , and brpT . (B) ), the number of overlapped genes with differential expression (fold change ≥ 1.5 or ≤ 0.667 and p -value ≤ 0.05) in Δ brpL and Δ ciaR was shown by Venn diagram. (C) The expression of genes in WT, Δ brpL and Δ brpL Δ ciaR was tested by qRT-PCR. All of the data were relative to their WT controls. (D) The biofilm biomass of strains was measured by CV staining. ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, Student’s t- test. Means and standard deviations from triplicate experiments are shown.

Techniques Used: Quantitative RT-PCR, Expressing, Staining

10) Product Images from "Role of Urease Enzymes in Stability of a 10-Species Oral Biofilm Consortium Cultivated in a Constant-Depth Film Fermenter"

Article Title: Role of Urease Enzymes in Stability of a 10-Species Oral Biofilm Consortium Cultivated in a Constant-Depth Film Fermenter

Journal: Infection and Immunity

doi: 10.1128/IAI.71.12.7188-7192.2003

Wild-type biofilms formed in the presence of urea. Biofilms were cultivated in complete BMMUG medium for the entire 11-day period. Organisms were recovered from the CDFF and enumerated by viable counting on selective media as detailed in the text. Viable counts are expressed as the average of the log CFU obtained per pocket, which provides the number of organisms that could be recovered from each of the five recessed pockets in the PTFE pans. Error bars indicate standard deviation. The data represent results from at least two complete runs of 11 days, and all platings were done in triplicate using three separate PTFE pans.
Figure Legend Snippet: Wild-type biofilms formed in the presence of urea. Biofilms were cultivated in complete BMMUG medium for the entire 11-day period. Organisms were recovered from the CDFF and enumerated by viable counting on selective media as detailed in the text. Viable counts are expressed as the average of the log CFU obtained per pocket, which provides the number of organisms that could be recovered from each of the five recessed pockets in the PTFE pans. Error bars indicate standard deviation. The data represent results from at least two complete runs of 11 days, and all platings were done in triplicate using three separate PTFE pans.

Techniques Used: Standard Deviation

11) Product Images from "Transcriptional Regulation of icaADBC by both IcaR and TcaR in Staphylococcus epidermidis"

Article Title: Transcriptional Regulation of icaADBC by both IcaR and TcaR in Staphylococcus epidermidis

Journal: Journal of Bacteriology

doi: 10.1128/JB.00524-18

icaA transcript, PIA synthesis, and biofilm production in 1457 mutants that produce less biofilm. (A and B) Northern blot analyses detecting icaA and icaR transcript in 1457 biofilm mutants in addition to PIA synthesis (A) and biofilm formation (B) compared to 1457 wild type (WT). Statistical analysis was performed using one-way ANOVA with Tukey’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. An RNA gel is shown as a loading control. PIA dot blot and biofilm analyses were assessed from three biological replicates.
Figure Legend Snippet: icaA transcript, PIA synthesis, and biofilm production in 1457 mutants that produce less biofilm. (A and B) Northern blot analyses detecting icaA and icaR transcript in 1457 biofilm mutants in addition to PIA synthesis (A) and biofilm formation (B) compared to 1457 wild type (WT). Statistical analysis was performed using one-way ANOVA with Tukey’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. An RNA gel is shown as a loading control. PIA dot blot and biofilm analyses were assessed from three biological replicates.

Techniques Used: Northern Blot, Dot Blot

12) Product Images from "Paradoxical antifungal activity and structural observations in biofilms formed by echinocandin-resistant Candida albicans clinical isolates"

Article Title: Paradoxical antifungal activity and structural observations in biofilms formed by echinocandin-resistant Candida albicans clinical isolates

Journal: Medical mycology

doi: 10.1093/mmy/myt007

Assessment of biofilm mass of Candida albicans reference strains and fks1 clinical isolates. Biofilms were grown in triplicate at a concentration of 1 × 10 6 cells/ml in buffered RPMI-1640 at 37°C for 24 h. Biofilm mass was quantified using the crystal violet assay. Light absorbance was measured in a plate reader at OD 630 nm. Each experiment was performed independently three times.
Figure Legend Snippet: Assessment of biofilm mass of Candida albicans reference strains and fks1 clinical isolates. Biofilms were grown in triplicate at a concentration of 1 × 10 6 cells/ml in buffered RPMI-1640 at 37°C for 24 h. Biofilm mass was quantified using the crystal violet assay. Light absorbance was measured in a plate reader at OD 630 nm. Each experiment was performed independently three times.

Techniques Used: Concentration Assay, Crystal Violet Assay

Ultrastructural assessment of biofilm morphology using scanning electron microscopy. (A) Scanning electron microscopy of representative Candida albicans fks1 mutants that form poor (4254), moderate (42286), and strong (53264) biofilms compared with reference strain SC5314. (B) Scanning electron microscopy view of pit-like cell surface structures identified on select C. albicans fks1 mutants.
Figure Legend Snippet: Ultrastructural assessment of biofilm morphology using scanning electron microscopy. (A) Scanning electron microscopy of representative Candida albicans fks1 mutants that form poor (4254), moderate (42286), and strong (53264) biofilms compared with reference strain SC5314. (B) Scanning electron microscopy view of pit-like cell surface structures identified on select C. albicans fks1 mutants.

Techniques Used: Electron Microscopy

Assessment of biofilm metabolic activity of Candida albicans reference strains and fks1 clinical isolates. Biofilms were grown in quadruplicate at a concentration of 1 × 10 6 cells/ml in buffered RPMI-1640 at 37° C for 24 h. The XTT assay was used to assay sessile metabolic activity. Formation of the colored formazan was subsequently measured at OD 490 nm. Each experiment was performed independently three times.
Figure Legend Snippet: Assessment of biofilm metabolic activity of Candida albicans reference strains and fks1 clinical isolates. Biofilms were grown in quadruplicate at a concentration of 1 × 10 6 cells/ml in buffered RPMI-1640 at 37° C for 24 h. The XTT assay was used to assay sessile metabolic activity. Formation of the colored formazan was subsequently measured at OD 490 nm. Each experiment was performed independently three times.

Techniques Used: Activity Assay, Concentration Assay, XTT Assay

13) Product Images from "Prevention of Staphylococcus aureus biofilm formation by antibiotics in 96-Microtiter Well Plates and Drip Flow Reactors: critical factors influencing outcomes"

Article Title: Prevention of Staphylococcus aureus biofilm formation by antibiotics in 96-Microtiter Well Plates and Drip Flow Reactors: critical factors influencing outcomes

Journal: Scientific Reports

doi: 10.1038/srep43854

Confocal laser scanning microscopy images of S. aureus ATCC 25923 biofilms grown in MWP ( A – C ) and DFR ( D – F ). ( A – C ) are representatives taken from different locations of one well in a MWP and D-F from different locations within one coupon in DFR. Biofilms are stained with LIVE/DEAD BacLight kit. Scale bars correspond to 30 μM.
Figure Legend Snippet: Confocal laser scanning microscopy images of S. aureus ATCC 25923 biofilms grown in MWP ( A – C ) and DFR ( D – F ). ( A – C ) are representatives taken from different locations of one well in a MWP and D-F from different locations within one coupon in DFR. Biofilms are stained with LIVE/DEAD BacLight kit. Scale bars correspond to 30 μM.

Techniques Used: Confocal Laser Scanning Microscopy, Staining

14) Product Images from "Mechanism of Fluconazole Resistance in Candida albicans Biofilms: Phase-Specific Role of Efflux Pumps and Membrane Sterols "

Article Title: Mechanism of Fluconazole Resistance in Candida albicans Biofilms: Phase-Specific Role of Efflux Pumps and Membrane Sterols

Journal: Infection and Immunity

doi: 10.1128/IAI.71.8.4333-4340.2003

Expression of CDR and MDR1 genes in (P) planktonic (P) and biofilm (B) forms of C. albicans . Total RNA was isolated from biofilms and planktonic cells grown for 6, 12, and 48 h. Sixty micrograms of total RNA was analyzed by Northern blotting using a CDR - or MDR1 -specific probe as described in Materials and Methods. Top, CDR transcript; middle, MDR1 transcript; bottom, 25S rRNA (loading control). Results are representative of three separate experiments.
Figure Legend Snippet: Expression of CDR and MDR1 genes in (P) planktonic (P) and biofilm (B) forms of C. albicans . Total RNA was isolated from biofilms and planktonic cells grown for 6, 12, and 48 h. Sixty micrograms of total RNA was analyzed by Northern blotting using a CDR - or MDR1 -specific probe as described in Materials and Methods. Top, CDR transcript; middle, MDR1 transcript; bottom, 25S rRNA (loading control). Results are representative of three separate experiments.

Techniques Used: Expressing, Isolation, Northern Blot

Percent growth inhibition of C. albicans biofilms exposed to high concentration of fluconazole. Percentages of inhibition for biofilms grown to the early (6 h), intermediate (12 h), or late (48 h) phase of development and exposed to 256 μg of fluconazole/ml were determined. Strains used were: CAF2-1 (wild type), DSY448 (Δ cdr1 ), DSY465 (Δ mdr1 ), DSY654 (Δ cdr1 Δ cdr2 ), and DSY1050 (Δ cdr1 Δ cdr2 Δ mdr1 ). For each strain, drug susceptibility decreased from the early to late phase of biofilm development. Additionally, deletion of two and three efflux pumps led to progressively decreasing susceptibility to fluconazole. Metabolic activity was normalized to the control without fluconazole, which was taken as 100%. Data (means ± standard deviations) are representative of three separate experiments. ∗, the wild-type (CAF2-1) strain showed 0% inhibition at all the time points.
Figure Legend Snippet: Percent growth inhibition of C. albicans biofilms exposed to high concentration of fluconazole. Percentages of inhibition for biofilms grown to the early (6 h), intermediate (12 h), or late (48 h) phase of development and exposed to 256 μg of fluconazole/ml were determined. Strains used were: CAF2-1 (wild type), DSY448 (Δ cdr1 ), DSY465 (Δ mdr1 ), DSY654 (Δ cdr1 Δ cdr2 ), and DSY1050 (Δ cdr1 Δ cdr2 Δ mdr1 ). For each strain, drug susceptibility decreased from the early to late phase of biofilm development. Additionally, deletion of two and three efflux pumps led to progressively decreasing susceptibility to fluconazole. Metabolic activity was normalized to the control without fluconazole, which was taken as 100%. Data (means ± standard deviations) are representative of three separate experiments. ∗, the wild-type (CAF2-1) strain showed 0% inhibition at all the time points.

Techniques Used: Inhibition, Concentration Assay, Activity Assay

Rh123 accumulation by early-, intermediate-, and mature-phase biofilms and planktonic cells of C. albicans. Data were analyzed by two-way analysis of variance, and a value of P
Figure Legend Snippet: Rh123 accumulation by early-, intermediate-, and mature-phase biofilms and planktonic cells of C. albicans. Data were analyzed by two-way analysis of variance, and a value of P

Techniques Used:

Variations in sterol profiles of C. albicans biofilms at different developmental phases. Sterol patterns for biofilms grown to the early (A), intermediate (B), or mature (C) phase were determined by GLC. (D) Percentages of sterols identified in C. albicans biofilms and planktonic cells (chromatograph not shown), determined from the corresponding peak areas and retention times relative to ergosterol. Peaks 1 to 7 (A to C) represent sterols described in panel D. SD, standard deviation.
Figure Legend Snippet: Variations in sterol profiles of C. albicans biofilms at different developmental phases. Sterol patterns for biofilms grown to the early (A), intermediate (B), or mature (C) phase were determined by GLC. (D) Percentages of sterols identified in C. albicans biofilms and planktonic cells (chromatograph not shown), determined from the corresponding peak areas and retention times relative to ergosterol. Peaks 1 to 7 (A to C) represent sterols described in panel D. SD, standard deviation.

Techniques Used: Gas Chromatography, Standard Deviation

15) Product Images from "Heparin Stimulates Staphylococcus aureus Biofilm Formation "

Article Title: Heparin Stimulates Staphylococcus aureus Biofilm Formation

Journal:

doi: 10.1128/IAI.73.8.4596-4606.2005

Effect of heparin on known biofilm formation mutants. A. Mutants (gene names noted) were assessed for biofilm formation in the microtiter dish assay (8-h biofilms) with heparin at 1,000 U/ml (shaded bars) or with saline (white bars). B. Relative PIA levels
Figure Legend Snippet: Effect of heparin on known biofilm formation mutants. A. Mutants (gene names noted) were assessed for biofilm formation in the microtiter dish assay (8-h biofilms) with heparin at 1,000 U/ml (shaded bars) or with saline (white bars). B. Relative PIA levels

Techniques Used:

Sodium heparin enhances S. aureus biofilm formation. The effect of heparin on the formation of S. aureus MZ100 biofilms on abiotic surfaces was assessed microscopically. Scanning electron micrographs of 12-hour-old S. aureus biofilms on polyvinylchloride
Figure Legend Snippet: Sodium heparin enhances S. aureus biofilm formation. The effect of heparin on the formation of S. aureus MZ100 biofilms on abiotic surfaces was assessed microscopically. Scanning electron micrographs of 12-hour-old S. aureus biofilms on polyvinylchloride

Techniques Used:

Sodium heparin increases the adherence of S. aureus to polystyrene. A. Dose response. Serial dilutions of sodium heparin were added to cultures and biofilms were allowed to form for 16 h, at which time nonadherent cells were removed by vigorous washing.
Figure Legend Snippet: Sodium heparin increases the adherence of S. aureus to polystyrene. A. Dose response. Serial dilutions of sodium heparin were added to cultures and biofilms were allowed to form for 16 h, at which time nonadherent cells were removed by vigorous washing.

Techniques Used:

Sodium heparin enhances S. aureus biofilm formation. The effect of heparin formation of S. aureus (MZ100) biofilms on abiotic surfaces was assessed microscopically. S. aureus biofilms (4 hours) formed on polystyrene were viewed with phase 2 microscopy
Figure Legend Snippet: Sodium heparin enhances S. aureus biofilm formation. The effect of heparin formation of S. aureus (MZ100) biofilms on abiotic surfaces was assessed microscopically. S. aureus biofilms (4 hours) formed on polystyrene were viewed with phase 2 microscopy

Techniques Used: Microscopy

16) Product Images from "Viable Compositional Analysis of an Eleven Species Oral Polymicrobial Biofilm"

Article Title: Viable Compositional Analysis of an Eleven Species Oral Polymicrobial Biofilm

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2016.00912

Daily combinational treatment impacts biofilm architecture by reducing total biomass . Multispecies biofilms were grown on PMMA for 7 days, as previously described. Following biofilm development, discs were washed and treated with B + DC daily for 5 days or brushed daily with intermittent cleansing on day 1 and 5. Untreated biofilms were also included for comparison. Biofilms were then processed and viewed on a JEOL-JSM 6400 scanning electron microscope and images assembled using Photoshop software. All images are shown at 2000 × magnifications and are representative of the sample. Scale bars represent 10 μm. Note the pores within the PMMA as denoted by arrows.
Figure Legend Snippet: Daily combinational treatment impacts biofilm architecture by reducing total biomass . Multispecies biofilms were grown on PMMA for 7 days, as previously described. Following biofilm development, discs were washed and treated with B + DC daily for 5 days or brushed daily with intermittent cleansing on day 1 and 5. Untreated biofilms were also included for comparison. Biofilms were then processed and viewed on a JEOL-JSM 6400 scanning electron microscope and images assembled using Photoshop software. All images are shown at 2000 × magnifications and are representative of the sample. Scale bars represent 10 μm. Note the pores within the PMMA as denoted by arrows.

Techniques Used: Microscopy, Software

Multi-species biofilm viability is greatly impacted by combinational treatment compared to monotherapy . Multispecies biofilms were grown on PMMA for 7 days, as previously described. Following maturation, biofilms were washed and either treated with a denture cleanser (DC) for 3 min, brushed only (B) , exposed to a combinational treatment of brushing before denture cleansing (B + DC) or brushing after denture cleansing (DC + B). Viability of total aerobes (A) , anaerobes (B) and Candida (C) was assessed by CFU counts. Untreated (UT) controls were also included. All testing was carried out in triplicate and on three independent occasions. Data represents mean ± SD, statistical analysis of treatments were compared to the untreated control ( *** p
Figure Legend Snippet: Multi-species biofilm viability is greatly impacted by combinational treatment compared to monotherapy . Multispecies biofilms were grown on PMMA for 7 days, as previously described. Following maturation, biofilms were washed and either treated with a denture cleanser (DC) for 3 min, brushed only (B) , exposed to a combinational treatment of brushing before denture cleansing (B + DC) or brushing after denture cleansing (DC + B). Viability of total aerobes (A) , anaerobes (B) and Candida (C) was assessed by CFU counts. Untreated (UT) controls were also included. All testing was carried out in triplicate and on three independent occasions. Data represents mean ± SD, statistical analysis of treatments were compared to the untreated control ( *** p

Techniques Used:

Biofilm compositional analysis of denture biofilms following oral hygiene regimens . Multispecies biofilms were grown on PMMA for 7 days before treated with the four therapies; denture-cleansing (DC), brushing (B), cleansing then brushing (DC + B) and brushing then cleansing (B + DC). Following treatment, each disc was sonicated before 50 μM of PMA was added and exposed to a 650 w halogen light source for 5 min to allow photo activation. Samples containing no PMA were also included to account for total biomass. DNA was extracted from each sample using the Qiagen DNA extraction kit, for quantification of each species using SYBR® GreenER™ based qPCR to determine the number of total and live cells remaining following treatment (A) . The composition of the biofilms following combinational treatment was also determined using species-specific primers (B) with total (i) and live (ii) cells shown. All testing was carried out in triplicate and on three independent occasions. Data represents mean ± SD, statistical analysis of treatments was compared to their respective untreated controls, in addition to total vs. live for each therapy ( * / # p
Figure Legend Snippet: Biofilm compositional analysis of denture biofilms following oral hygiene regimens . Multispecies biofilms were grown on PMMA for 7 days before treated with the four therapies; denture-cleansing (DC), brushing (B), cleansing then brushing (DC + B) and brushing then cleansing (B + DC). Following treatment, each disc was sonicated before 50 μM of PMA was added and exposed to a 650 w halogen light source for 5 min to allow photo activation. Samples containing no PMA were also included to account for total biomass. DNA was extracted from each sample using the Qiagen DNA extraction kit, for quantification of each species using SYBR® GreenER™ based qPCR to determine the number of total and live cells remaining following treatment (A) . The composition of the biofilms following combinational treatment was also determined using species-specific primers (B) with total (i) and live (ii) cells shown. All testing was carried out in triplicate and on three independent occasions. Data represents mean ± SD, statistical analysis of treatments was compared to their respective untreated controls, in addition to total vs. live for each therapy ( * / # p

Techniques Used: Sonication, Activation Assay, DNA Extraction, Real-time Polymerase Chain Reaction

Daily cleaning of denture biofilms reduces the biofilm biomass and viability . Multispecies complex biofilms were grown on PMMA for 7 days, as previously described. Following maturation, biofilms were washed and either treated daily with brushing and denture cleansing (B + DC) or brushed daily with the addition of a DC on day 1 and 5 only (B) . Viability of total aerobes (Ai) , anaerobes (Aii) , and Candida (Aiii) was assessed by CFU. Biofilms were also treated with PMA and exposed to a 650 w halogen light for live-dead PCR analysis. Samples containing no PMA were also included to account for total biomass. DNA was extracted from each sample using the Qiagen DNA extraction kit, for quantification of total (Bi) and live (Bii) bacteria and total (Biii) and live yeast (Biv) using SYBR® GreenER™ based qPCR. All testing was carried out in triplicate and on three independent occasions. Data represents mean ± SD, statistical analysis of treatments was compared to their respective untreated controls, in addition to total vs. live for each therapy ( * p
Figure Legend Snippet: Daily cleaning of denture biofilms reduces the biofilm biomass and viability . Multispecies complex biofilms were grown on PMMA for 7 days, as previously described. Following maturation, biofilms were washed and either treated daily with brushing and denture cleansing (B + DC) or brushed daily with the addition of a DC on day 1 and 5 only (B) . Viability of total aerobes (Ai) , anaerobes (Aii) , and Candida (Aiii) was assessed by CFU. Biofilms were also treated with PMA and exposed to a 650 w halogen light for live-dead PCR analysis. Samples containing no PMA were also included to account for total biomass. DNA was extracted from each sample using the Qiagen DNA extraction kit, for quantification of total (Bi) and live (Bii) bacteria and total (Biii) and live yeast (Biv) using SYBR® GreenER™ based qPCR. All testing was carried out in triplicate and on three independent occasions. Data represents mean ± SD, statistical analysis of treatments was compared to their respective untreated controls, in addition to total vs. live for each therapy ( * p

Techniques Used: Polymerase Chain Reaction, DNA Extraction, Real-time Polymerase Chain Reaction

Live cells imaging reveals viable cells within pores following treatment . Multispecies biofilms were grown on PMMA for 7 days, as previously described. Following biofilm development, discs were washed and treated with B + DC daily for 5 days or brushed daily with intermittent cleansing on day 1 and 5. Untreated biofilms were also included for comparison. Images were stained with SYTO9 and PI to show live and dead cells remaining following treatment and viewed under a CLSM (Leica SP5). All images are shown at 20 × magnification and scale bars represent 20 μm. Note the pores within the PMMA as denoted by the arrows.
Figure Legend Snippet: Live cells imaging reveals viable cells within pores following treatment . Multispecies biofilms were grown on PMMA for 7 days, as previously described. Following biofilm development, discs were washed and treated with B + DC daily for 5 days or brushed daily with intermittent cleansing on day 1 and 5. Untreated biofilms were also included for comparison. Images were stained with SYTO9 and PI to show live and dead cells remaining following treatment and viewed under a CLSM (Leica SP5). All images are shown at 20 × magnification and scale bars represent 20 μm. Note the pores within the PMMA as denoted by the arrows.

Techniques Used: Imaging, Staining, Confocal Laser Scanning Microscopy

Sequential treatment of denture biofilm protocol . PMMA discs were placed in 24 well plates for biofilm culture. Biofilms were treated daily with brushing and a denture cleanser for 5 days or were brushed every day with denture cleansing on day 1 and day 5. Untreated controls were maintained in artificial saliva during treatments.
Figure Legend Snippet: Sequential treatment of denture biofilm protocol . PMMA discs were placed in 24 well plates for biofilm culture. Biofilms were treated daily with brushing and a denture cleanser for 5 days or were brushed every day with denture cleansing on day 1 and day 5. Untreated controls were maintained in artificial saliva during treatments.

Techniques Used:

17) Product Images from "Transcriptome Assembly and Profiling of Candida auris Reveals Novel Insights into Biofilm-Mediated Resistance"

Article Title: Transcriptome Assembly and Profiling of Candida auris Reveals Novel Insights into Biofilm-Mediated Resistance

Journal: mSphere

doi: 10.1128/mSphere.00334-18

Quality control and differential expression analysis of C. auris biofilms. Principal-component analysis displays the largest variance along PC1 (56%) and the second largest variance between samples along PC2 (15%) (A). Venn diagrams of the genes upregulated (B) and downregulated (C) in biofilm time points (4, 12, and 24 h) compared to 0 h.
Figure Legend Snippet: Quality control and differential expression analysis of C. auris biofilms. Principal-component analysis displays the largest variance along PC1 (56%) and the second largest variance between samples along PC2 (15%) (A). Venn diagrams of the genes upregulated (B) and downregulated (C) in biofilm time points (4, 12, and 24 h) compared to 0 h.

Techniques Used: Expressing

Formation and development of Candida auris biofilms. Schematic representation of the transcriptional mediators of the three main stages of C. auris biofilm development: adherence of yeast cells to surface (early phase), proliferation (intermediate phase), and maturation into a structured biofilm (mature phase).
Figure Legend Snippet: Formation and development of Candida auris biofilms. Schematic representation of the transcriptional mediators of the three main stages of C. auris biofilm development: adherence of yeast cells to surface (early phase), proliferation (intermediate phase), and maturation into a structured biofilm (mature phase).

Techniques Used:

Functional annotation of differentially expressed genes reveals upregulation of drug transporters. Gene distribution of significantly upregulated C. auris genes in 24-h biofilms relative to planktonic cells, grouped into biological process (BP), cellular component (CC), and metabolic function (MF) gene ontology categories (A). Log 2 fold change of upregulated ABC and MFS drug transporters within 24-h biofilms (B). All GO terms have a P value of
Figure Legend Snippet: Functional annotation of differentially expressed genes reveals upregulation of drug transporters. Gene distribution of significantly upregulated C. auris genes in 24-h biofilms relative to planktonic cells, grouped into biological process (BP), cellular component (CC), and metabolic function (MF) gene ontology categories (A). Log 2 fold change of upregulated ABC and MFS drug transporters within 24-h biofilms (B). All GO terms have a P value of

Techniques Used: Functional Assay

Efflux pump activity is increased in Candida auris biofilms. Candida auris biofilms were grown for 4, 12, and 24 h in black-bottomed 96-well plates. In addition, planktonic cells were standardized to 5 × 10 7 cells/ml, all cells were incubated with 100 µg/ml of Ala-Nap, and fluorescence measurements were read at 30-s intervals over 60 min (excitation, 355 nm; emission, 460 nm). Data represent the mean + standard deviation of 4 isolates repeated on 3 independent occasions. Data presented are relative fluorescence units (RFU) normalized per individual cell. *, P
Figure Legend Snippet: Efflux pump activity is increased in Candida auris biofilms. Candida auris biofilms were grown for 4, 12, and 24 h in black-bottomed 96-well plates. In addition, planktonic cells were standardized to 5 × 10 7 cells/ml, all cells were incubated with 100 µg/ml of Ala-Nap, and fluorescence measurements were read at 30-s intervals over 60 min (excitation, 355 nm; emission, 460 nm). Data represent the mean + standard deviation of 4 isolates repeated on 3 independent occasions. Data presented are relative fluorescence units (RFU) normalized per individual cell. *, P

Techniques Used: Activity Assay, Incubation, Fluorescence, Standard Deviation

Candida auris biofilm development correlates with increased antifungal tolerance. Candida auris biofilms were standardized at 1 × 10 6 CFU/ml and grown for 4, 12, and 24 h. Biofilm biomass was then quantified using the crystal violet assay, with the composition of biofilm cells enumerated using qPCR and represented by a box-and-whisker plot as the total biomass of four C. auris isolates (A, left y axis). Planktonic susceptibility testing was performed against serially diluted miconazole, micafungin, and amphotericin B concentrations using the CLSI guidelines, with biofilm susceptibility testing performed using the XTT assay and with median MIC values plotted (A, right y axis). In addition, biofilms were grown, fixed, and processed for SEM before imaging using a JEOL-JSM-6400 scanning electron microscope. Micrographs represent lower magnification (×1,000) and higher magnification (inset, ×5,000) of biofilms grown for 4 h (Bi), 12 h (Bii), and 24 h (Biii).
Figure Legend Snippet: Candida auris biofilm development correlates with increased antifungal tolerance. Candida auris biofilms were standardized at 1 × 10 6 CFU/ml and grown for 4, 12, and 24 h. Biofilm biomass was then quantified using the crystal violet assay, with the composition of biofilm cells enumerated using qPCR and represented by a box-and-whisker plot as the total biomass of four C. auris isolates (A, left y axis). Planktonic susceptibility testing was performed against serially diluted miconazole, micafungin, and amphotericin B concentrations using the CLSI guidelines, with biofilm susceptibility testing performed using the XTT assay and with median MIC values plotted (A, right y axis). In addition, biofilms were grown, fixed, and processed for SEM before imaging using a JEOL-JSM-6400 scanning electron microscope. Micrographs represent lower magnification (×1,000) and higher magnification (inset, ×5,000) of biofilms grown for 4 h (Bi), 12 h (Bii), and 24 h (Biii).

Techniques Used: Crystal Violet Assay, Real-time Polymerase Chain Reaction, Whisker Assay, XTT Assay, Imaging, Microscopy

18) Product Images from "Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions"

Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.00258

Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. Biofilms were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p
Figure Legend Snippet: Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. Biofilms were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p

Techniques Used: Sonication, Polymerase Chain Reaction

Candida albicans mycofilms facilitates Staphylococcus aureus biofilm formation. Mono- and dual-species biofilms were standardized 1 × 10 6 CFU/mL and grown on 96-well plates for 90 min, 6 and 24 h. Biofilms were then washed with phosphate buffered saline (PBS) and biomass assessed using the crystal violet assay (A). Standardized biofilms were grown before being sonicated to remove the biomass. Live/dead PCR was then used to extract DNA and determine total and live colony forming equivalents (CFE) from mono- and dual-species biofilms (B) . Biofilm morphology was then analyzed using CSLM. Biofilms were grown before being fluorescently stained using calcofluor white and SYTO9 ® dyes. Resulting biofilms were then viewed on a Leica SP5 laser scanning confocal microscope and images were then processed and analyzed using Volocity 3D Image Analysis Software (C) . Results represent data from three independent occasions. Statistical analysis compares dual-species biofilms to their mono-species equivalent ( ∗ p
Figure Legend Snippet: Candida albicans mycofilms facilitates Staphylococcus aureus biofilm formation. Mono- and dual-species biofilms were standardized 1 × 10 6 CFU/mL and grown on 96-well plates for 90 min, 6 and 24 h. Biofilms were then washed with phosphate buffered saline (PBS) and biomass assessed using the crystal violet assay (A). Standardized biofilms were grown before being sonicated to remove the biomass. Live/dead PCR was then used to extract DNA and determine total and live colony forming equivalents (CFE) from mono- and dual-species biofilms (B) . Biofilm morphology was then analyzed using CSLM. Biofilms were grown before being fluorescently stained using calcofluor white and SYTO9 ® dyes. Resulting biofilms were then viewed on a Leica SP5 laser scanning confocal microscope and images were then processed and analyzed using Volocity 3D Image Analysis Software (C) . Results represent data from three independent occasions. Statistical analysis compares dual-species biofilms to their mono-species equivalent ( ∗ p

Techniques Used: Crystal Violet Assay, Sonication, Polymerase Chain Reaction, Staining, Microscopy, Software

Scanning electron micrograph of S. aureus colonizing C. albicans hyphae within dual-species biofilms. Dual-species biofilms were grown for 24 h before being fixed, processed, and imaged using a JEOL-JSM 6400 scanning electron microscope. S. aureus colonies can be seen adhering and embedded within the hyphal meshwork of C. albicans . White arrows indicate clusters of S. aureus colonies encased within extracellular matrix (ECM). Scale bar represents 5 μm at × 5000 magnification.
Figure Legend Snippet: Scanning electron micrograph of S. aureus colonizing C. albicans hyphae within dual-species biofilms. Dual-species biofilms were grown for 24 h before being fixed, processed, and imaged using a JEOL-JSM 6400 scanning electron microscope. S. aureus colonies can be seen adhering and embedded within the hyphal meshwork of C. albicans . White arrows indicate clusters of S. aureus colonies encased within extracellular matrix (ECM). Scale bar represents 5 μm at × 5000 magnification.

Techniques Used: Microscopy

Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p
Figure Legend Snippet: Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p

Techniques Used: SYBR Green Assay, Fluorescence, Real-time Polymerase Chain Reaction, Staining, Incubation

19) Product Images from "Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions"

Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.00258

Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. Biofilms were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p
Figure Legend Snippet: Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. Biofilms were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p

Techniques Used: Sonication, Polymerase Chain Reaction

Candida albicans mycofilms facilitates Staphylococcus aureus biofilm formation. Mono- and dual-species biofilms were standardized 1 × 10 6 CFU/mL and grown on 96-well plates for 90 min, 6 and 24 h. Biofilms were then washed with phosphate buffered saline (PBS) and biomass assessed using the crystal violet assay (A). Standardized biofilms were grown before being sonicated to remove the biomass. Live/dead PCR was then used to extract DNA and determine total and live colony forming equivalents (CFE) from mono- and dual-species biofilms (B) . Biofilm morphology was then analyzed using CSLM. Biofilms were grown before being fluorescently stained using calcofluor white and SYTO9 ® dyes. Resulting biofilms were then viewed on a Leica SP5 laser scanning confocal microscope and images were then processed and analyzed using Volocity 3D Image Analysis Software (C) . Results represent data from three independent occasions. Statistical analysis compares dual-species biofilms to their mono-species equivalent ( ∗ p
Figure Legend Snippet: Candida albicans mycofilms facilitates Staphylococcus aureus biofilm formation. Mono- and dual-species biofilms were standardized 1 × 10 6 CFU/mL and grown on 96-well plates for 90 min, 6 and 24 h. Biofilms were then washed with phosphate buffered saline (PBS) and biomass assessed using the crystal violet assay (A). Standardized biofilms were grown before being sonicated to remove the biomass. Live/dead PCR was then used to extract DNA and determine total and live colony forming equivalents (CFE) from mono- and dual-species biofilms (B) . Biofilm morphology was then analyzed using CSLM. Biofilms were grown before being fluorescently stained using calcofluor white and SYTO9 ® dyes. Resulting biofilms were then viewed on a Leica SP5 laser scanning confocal microscope and images were then processed and analyzed using Volocity 3D Image Analysis Software (C) . Results represent data from three independent occasions. Statistical analysis compares dual-species biofilms to their mono-species equivalent ( ∗ p

Techniques Used: Crystal Violet Assay, Sonication, Polymerase Chain Reaction, Staining, Microscopy, Software

Scanning electron micrograph of S. aureus colonizing C. albicans hyphae within dual-species biofilms. Dual-species biofilms were grown for 24 h before being fixed, processed, and imaged using a JEOL-JSM 6400 scanning electron microscope. S. aureus colonies can be seen adhering and embedded within the hyphal meshwork of C. albicans . White arrows indicate clusters of S. aureus colonies encased within extracellular matrix (ECM). Scale bar represents 5 μm at × 5000 magnification.
Figure Legend Snippet: Scanning electron micrograph of S. aureus colonizing C. albicans hyphae within dual-species biofilms. Dual-species biofilms were grown for 24 h before being fixed, processed, and imaged using a JEOL-JSM 6400 scanning electron microscope. S. aureus colonies can be seen adhering and embedded within the hyphal meshwork of C. albicans . White arrows indicate clusters of S. aureus colonies encased within extracellular matrix (ECM). Scale bar represents 5 μm at × 5000 magnification.

Techniques Used: Microscopy

Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p
Figure Legend Snippet: Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p

Techniques Used: SYBR Green Assay, Fluorescence, Real-time Polymerase Chain Reaction, Staining, Incubation

20) Product Images from "In vitro characterization of biofilms formed by Kingella kingae"

Article Title: In vitro characterization of biofilms formed by Kingella kingae

Journal: Molecular oral microbiology

doi: 10.1111/omi.12176

Transcript levels of genes pilA1, pilA2, fimB for type IV pili production in biofilm-forming strains PYKK081, PYKK109, 0211+12480 and non-biofilm forming strains ATCC 23330, 0604+15110. Transcripts levels are expressed relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH), SEM≥5%.
Figure Legend Snippet: Transcript levels of genes pilA1, pilA2, fimB for type IV pili production in biofilm-forming strains PYKK081, PYKK109, 0211+12480 and non-biofilm forming strains ATCC 23330, 0604+15110. Transcripts levels are expressed relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH), SEM≥5%.

Techniques Used:

Biofilm formation by 58 corroding and 21 noncorroding K. kingae strains in 96-well microtiter plates. Biofilm was quantitated using a crystal violet binding assay. Absorbance at 595 nm is proportional to biofilm biomass.
Figure Legend Snippet: Biofilm formation by 58 corroding and 21 noncorroding K. kingae strains in 96-well microtiter plates. Biofilm was quantitated using a crystal violet binding assay. Absorbance at 595 nm is proportional to biofilm biomass.

Techniques Used: Crystal Violet Binding Assay

Colony morphologies on agar and biofilm phenotypes in broth of K. kingae corroding strain PYKK109 (top panels) and noncorroding strain ATCC 23330 (bottom panels). From left to right: bacterial colonies on agar; the agar surface beneath the colony after the colony was removed by rinsing with water; biofilm phenotype in broth (side view); biofilm phenotype in broth (bottom view).
Figure Legend Snippet: Colony morphologies on agar and biofilm phenotypes in broth of K. kingae corroding strain PYKK109 (top panels) and noncorroding strain ATCC 23330 (bottom panels). From left to right: bacterial colonies on agar; the agar surface beneath the colony after the colony was removed by rinsing with water; biofilm phenotype in broth (side view); biofilm phenotype in broth (bottom view).

Techniques Used:

Effects of proteinase K and DNase I on K. kingae biofilm formation. In the left-hand graph, the enzymes were added to the broth at the time of inoculation and biofilm was quantitated after 24 h. In the right-hand graph, biofilms were cultured for 24 h and then treated with enzymes for 1 h. A 595 values are proportional to biofilm biomass. Asterisks indicate values significantly different from no-enzyme controls ( P
Figure Legend Snippet: Effects of proteinase K and DNase I on K. kingae biofilm formation. In the left-hand graph, the enzymes were added to the broth at the time of inoculation and biofilm was quantitated after 24 h. In the right-hand graph, biofilms were cultured for 24 h and then treated with enzymes for 1 h. A 595 values are proportional to biofilm biomass. Asterisks indicate values significantly different from no-enzyme controls ( P

Techniques Used: Cell Culture

Autoaggregation, biofilm formation, and agar corrosion by K. kingae wild-type strain PYKK109 and pilus mutant strain VS1001. (A) Autoaggregation of cells in microcentrifuge tubes. (B) Biofilm formation on glass slides. In panel B, biofilms were visualized by staining with the green fluorescent nucleic acid stain SYTO9. (C) AFM topographic image (top) and cross-sectional profile (bottom) of agar surface after the bacteria were washed from the agar surfaces. For the pitted sample, the free amplitude and setpoint were 156 nm and 83 nm, respectively. For the control sample, those values were 208 nm and 145 nm. Areas of agar depression are designated with white circles.
Figure Legend Snippet: Autoaggregation, biofilm formation, and agar corrosion by K. kingae wild-type strain PYKK109 and pilus mutant strain VS1001. (A) Autoaggregation of cells in microcentrifuge tubes. (B) Biofilm formation on glass slides. In panel B, biofilms were visualized by staining with the green fluorescent nucleic acid stain SYTO9. (C) AFM topographic image (top) and cross-sectional profile (bottom) of agar surface after the bacteria were washed from the agar surfaces. For the pitted sample, the free amplitude and setpoint were 156 nm and 83 nm, respectively. For the control sample, those values were 208 nm and 145 nm. Areas of agar depression are designated with white circles.

Techniques Used: Mutagenesis, Staining

21) Product Images from "Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions"

Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.00258

Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. Biofilms were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p
Figure Legend Snippet: Inter-kingdom interactions decrease Staphylococcus aureus sensitivity to miconazole. Biofilms were grown for 24 h on a cellulose matrix based hydrogel model before being washed with PBS and treated with 40 mg/L of miconazole for a further 24 h. After treatment, the cellulose matrix was removed and sonicated to dislodge the biofilm biomass. Live/dead PCR was then used to extract DNA and quantify total and live CFE. Data is presented as the CFE of live cells comparing treated mono- and dual-species biofilms. Data represents duplicate samples from three independent time points with significance achieved with ∗∗ p

Techniques Used: Sonication, Polymerase Chain Reaction

Candida albicans mycofilms facilitates Staphylococcus aureus biofilm formation. Mono- and dual-species biofilms were standardized 1 × 10 6 CFU/mL and grown on 96-well plates for 90 min, 6 and 24 h. Biofilms were then washed with phosphate buffered saline (PBS) and biomass assessed using the crystal violet assay (A). Standardized biofilms were grown before being sonicated to remove the biomass. Live/dead PCR was then used to extract DNA and determine total and live colony forming equivalents (CFE) from mono- and dual-species biofilms (B) . Biofilm morphology was then analyzed using CSLM. Biofilms were grown before being fluorescently stained using calcofluor white and SYTO9 ® dyes. Resulting biofilms were then viewed on a Leica SP5 laser scanning confocal microscope and images were then processed and analyzed using Volocity 3D Image Analysis Software (C) . Results represent data from three independent occasions. Statistical analysis compares dual-species biofilms to their mono-species equivalent ( ∗ p
Figure Legend Snippet: Candida albicans mycofilms facilitates Staphylococcus aureus biofilm formation. Mono- and dual-species biofilms were standardized 1 × 10 6 CFU/mL and grown on 96-well plates for 90 min, 6 and 24 h. Biofilms were then washed with phosphate buffered saline (PBS) and biomass assessed using the crystal violet assay (A). Standardized biofilms were grown before being sonicated to remove the biomass. Live/dead PCR was then used to extract DNA and determine total and live colony forming equivalents (CFE) from mono- and dual-species biofilms (B) . Biofilm morphology was then analyzed using CSLM. Biofilms were grown before being fluorescently stained using calcofluor white and SYTO9 ® dyes. Resulting biofilms were then viewed on a Leica SP5 laser scanning confocal microscope and images were then processed and analyzed using Volocity 3D Image Analysis Software (C) . Results represent data from three independent occasions. Statistical analysis compares dual-species biofilms to their mono-species equivalent ( ∗ p

Techniques Used: Crystal Violet Assay, Sonication, Polymerase Chain Reaction, Staining, Microscopy, Software

Scanning electron micrograph of S. aureus colonizing C. albicans hyphae within dual-species biofilms. Dual-species biofilms were grown for 24 h before being fixed, processed, and imaged using a JEOL-JSM 6400 scanning electron microscope. S. aureus colonies can be seen adhering and embedded within the hyphal meshwork of C. albicans . White arrows indicate clusters of S. aureus colonies encased within extracellular matrix (ECM). Scale bar represents 5 μm at × 5000 magnification.
Figure Legend Snippet: Scanning electron micrograph of S. aureus colonizing C. albicans hyphae within dual-species biofilms. Dual-species biofilms were grown for 24 h before being fixed, processed, and imaged using a JEOL-JSM 6400 scanning electron microscope. S. aureus colonies can be seen adhering and embedded within the hyphal meshwork of C. albicans . White arrows indicate clusters of S. aureus colonies encased within extracellular matrix (ECM). Scale bar represents 5 μm at × 5000 magnification.

Techniques Used: Microscopy

Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p
Figure Legend Snippet: Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p

Techniques Used: SYBR Green Assay, Fluorescence, Real-time Polymerase Chain Reaction, Staining, Incubation

22) Product Images from "The Relative Contributions of Physical Structure and Cell Density to the Antibiotic Susceptibility of Bacteria in Biofilms"

Article Title: The Relative Contributions of Physical Structure and Cell Density to the Antibiotic Susceptibility of Bacteria in Biofilms

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.06480-11

Survival of E. coli MG1655 csrA biofilm and planktonic cells in the presence of high concentrations of antibiotics. Cells were exposed to 40× MIC of the indicated drugs, and viable cell densities were estimated at 3 h. (A) Control, no drug; (B)
Figure Legend Snippet: Survival of E. coli MG1655 csrA biofilm and planktonic cells in the presence of high concentrations of antibiotics. Cells were exposed to 40× MIC of the indicated drugs, and viable cell densities were estimated at 3 h. (A) Control, no drug; (B)

Techniques Used:

Survival of S. aureus biofilms and planktonic cells in the presence of low concentrations of gentamicin. Cells were exposed to 10× MIC of gentamicin, and viable cell densities were estimated at 3 h. Bars represent the standard errors. Letters
Figure Legend Snippet: Survival of S. aureus biofilms and planktonic cells in the presence of low concentrations of gentamicin. Cells were exposed to 10× MIC of gentamicin, and viable cell densities were estimated at 3 h. Bars represent the standard errors. Letters

Techniques Used:

Survival of E. coli MG1655 csrA biofilm and planktonic cells in the presence of low concentrations of colistin and streptomycin. Cells were exposed to 10× MIC of colistin or streptomycin, and viable cell densities were estimated at 3 h. (A) Colistin;
Figure Legend Snippet: Survival of E. coli MG1655 csrA biofilm and planktonic cells in the presence of low concentrations of colistin and streptomycin. Cells were exposed to 10× MIC of colistin or streptomycin, and viable cell densities were estimated at 3 h. (A) Colistin;

Techniques Used:

Calibration experiments for biofilm growth and disruption. (A, B) Extent of disruption over time. Biofilms were grown and prepared as described for the time-kill experiments, followed by disruption for 0, 10, 20, or 30 s. White bars, no disruption; black
Figure Legend Snippet: Calibration experiments for biofilm growth and disruption. (A, B) Extent of disruption over time. Biofilms were grown and prepared as described for the time-kill experiments, followed by disruption for 0, 10, 20, or 30 s. White bars, no disruption; black

Techniques Used:

Survival of S. aureus 35556 biofilm and planktonic cells in the presence of high antibiotic concentrations. Cells were exposed to 40× MIC of the indicated drugs, and viable cell densities were estimated at 3 h. (A) Control, no drug; (B) ciprofloxacin;
Figure Legend Snippet: Survival of S. aureus 35556 biofilm and planktonic cells in the presence of high antibiotic concentrations. Cells were exposed to 40× MIC of the indicated drugs, and viable cell densities were estimated at 3 h. (A) Control, no drug; (B) ciprofloxacin;

Techniques Used:

Relative efficacy of antibiotics under different culture conditions. Relative efficacy is the ratio of the average relative kill of the given antibiotic in the given condition to the average relative kill of the antibiotic in intact biofilm conditions.
Figure Legend Snippet: Relative efficacy of antibiotics under different culture conditions. Relative efficacy is the ratio of the average relative kill of the given antibiotic in the given condition to the average relative kill of the antibiotic in intact biofilm conditions.

Techniques Used:

23) Product Images from "Rhesus θ-defensin-1 (RTD-1) exhibits in vitro and in vivo activity against cystic fibrosis strains of Pseudomonas aeruginosa"

Article Title: Rhesus θ-defensin-1 (RTD-1) exhibits in vitro and in vivo activity against cystic fibrosis strains of Pseudomonas aeruginosa

Journal: Journal of Antimicrobial Chemotherapy

doi: 10.1093/jac/dkv301

Activity of RTD-1 against PAO1 biofilms. Biofilms were grown on polycarbonate membranes placed on M63 agarose plates as described in the Materials and methods section. (a) Electron microscopy image of PAO1 aggregates on polycarbonate membranes. After
Figure Legend Snippet: Activity of RTD-1 against PAO1 biofilms. Biofilms were grown on polycarbonate membranes placed on M63 agarose plates as described in the Materials and methods section. (a) Electron microscopy image of PAO1 aggregates on polycarbonate membranes. After

Techniques Used: Activity Assay, Electron Microscopy

24) Product Images from "Rhesus θ-defensin-1 (RTD-1) exhibits in vitro and in vivo activity against cystic fibrosis strains of Pseudomonas aeruginosa"

Article Title: Rhesus θ-defensin-1 (RTD-1) exhibits in vitro and in vivo activity against cystic fibrosis strains of Pseudomonas aeruginosa

Journal: Journal of Antimicrobial Chemotherapy

doi: 10.1093/jac/dkv301

Activity of RTD-1 against PAO1 biofilms. Biofilms were grown on polycarbonate membranes placed on M63 agarose plates as described in the Materials and methods section. (a) Electron microscopy image of PAO1 aggregates on polycarbonate membranes. After
Figure Legend Snippet: Activity of RTD-1 against PAO1 biofilms. Biofilms were grown on polycarbonate membranes placed on M63 agarose plates as described in the Materials and methods section. (a) Electron microscopy image of PAO1 aggregates on polycarbonate membranes. After

Techniques Used: Activity Assay, Electron Microscopy

25) Product Images from "Evaluating Streptococcus mutans Strain Dependent Characteristics in a Polymicrobial Biofilm Community"

Article Title: Evaluating Streptococcus mutans Strain Dependent Characteristics in a Polymicrobial Biofilm Community

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.01498

Polymicrobial cariogenic biofilm biomass is influenced by carbohydrate source. (A) Polymicrobial caries biofilm model containing standardized L. casei , V. dispar , F. nucleatum , A. naeslundii and S. mutans isolates derived from either caries and caries-free patients grown at was grown in artificial saliva (AS) + either galactose or sucrose at 37°C in 5% CO 2 for 5 days, with spent supernatants being replaced with fresh AS every 24 h. Biomass crystal violet staining was quantified spectrophotometrically at 570 nm. Significant increase in growth compared to the 24 h culture ( ∗∗ p
Figure Legend Snippet: Polymicrobial cariogenic biofilm biomass is influenced by carbohydrate source. (A) Polymicrobial caries biofilm model containing standardized L. casei , V. dispar , F. nucleatum , A. naeslundii and S. mutans isolates derived from either caries and caries-free patients grown at was grown in artificial saliva (AS) + either galactose or sucrose at 37°C in 5% CO 2 for 5 days, with spent supernatants being replaced with fresh AS every 24 h. Biomass crystal violet staining was quantified spectrophotometrically at 570 nm. Significant increase in growth compared to the 24 h culture ( ∗∗ p

Techniques Used: Derivative Assay, Staining

Monospecies cariogenic biofilm biomass is influenced by carbohydrate source. Biomass was quantified spectrophotometrically by reading absorbance at 570 nm in a microtiter plate reader (FluoStar Omega, BMG Labtech). Four replicates were used for each isolate and was carried out on triplicate separate occasions, with the mean of each represented. Data represents mean with significance ∗ p
Figure Legend Snippet: Monospecies cariogenic biofilm biomass is influenced by carbohydrate source. Biomass was quantified spectrophotometrically by reading absorbance at 570 nm in a microtiter plate reader (FluoStar Omega, BMG Labtech). Four replicates were used for each isolate and was carried out on triplicate separate occasions, with the mean of each represented. Data represents mean with significance ∗ p

Techniques Used:

Polymicrobial cariogenic biofilm viability is influenced by carbohydrate source. (A) Polymicrobial caries biofilm model containing standardized L. casei , V. dispar , F. nucleatum , A. naeslundii and S. mutans isolates derived from either caries and caries-free patients grown at was grown in artificial saliva (AS) + galactose or sucrose at 37°C in 5% CO 2 for 5 days, with spent supernatants being replaced with fresh AS every 24 h. Metabolic activity (AlamarBlue) was quantified spectrophotometrically at 490 nm. Significant increase in growth compared to the 24 h culture ( ∗∗ p
Figure Legend Snippet: Polymicrobial cariogenic biofilm viability is influenced by carbohydrate source. (A) Polymicrobial caries biofilm model containing standardized L. casei , V. dispar , F. nucleatum , A. naeslundii and S. mutans isolates derived from either caries and caries-free patients grown at was grown in artificial saliva (AS) + galactose or sucrose at 37°C in 5% CO 2 for 5 days, with spent supernatants being replaced with fresh AS every 24 h. Metabolic activity (AlamarBlue) was quantified spectrophotometrically at 490 nm. Significant increase in growth compared to the 24 h culture ( ∗∗ p

Techniques Used: Derivative Assay, Activity Assay

26) Product Images from "Anti-caries DNA vaccine-induced secretory immunoglobulin A antibodies inhibit formation of Streptococcus mutans biofilms in vitro"

Article Title: Anti-caries DNA vaccine-induced secretory immunoglobulin A antibodies inhibit formation of Streptococcus mutans biofilms in vitro

Journal: Acta Pharmacologica Sinica

doi: 10.1038/aps.2012.145

Scanning electron microscopy (SEM) investigation. Surface of hydroxyapatite disks at 2000× (A) and 5000× (B) magnification (HA disk was a standard hexagonal structure). S. mutans 16 h biofilm at 8000× (C) and 20000× (D) magnification ( S. mutans biofilm uniformly covered the surface of the HA disks). The surface of the hydroxyapatite disks at 2000× (E) and 5000× (F) magnification after biofilm resuspension (few remaining bacteria were observed: arrows).
Figure Legend Snippet: Scanning electron microscopy (SEM) investigation. Surface of hydroxyapatite disks at 2000× (A) and 5000× (B) magnification (HA disk was a standard hexagonal structure). S. mutans 16 h biofilm at 8000× (C) and 20000× (D) magnification ( S. mutans biofilm uniformly covered the surface of the HA disks). The surface of the hydroxyapatite disks at 2000× (E) and 5000× (F) magnification after biofilm resuspension (few remaining bacteria were observed: arrows).

Techniques Used: Electron Microscopy

Effect of S-IgA pretreated HA disks on biofilm formation. Acquired pellicle formed by saliva from groups A and B. The pellicle formed by saliva from group A could be observed to have more S-IgA (green) on the HA disks than the pellicle formed by saliva from group B.
Figure Legend Snippet: Effect of S-IgA pretreated HA disks on biofilm formation. Acquired pellicle formed by saliva from groups A and B. The pellicle formed by saliva from group A could be observed to have more S-IgA (green) on the HA disks than the pellicle formed by saliva from group B.

Techniques Used:

27) Product Images from "Differential Gene Expression to Investigate the Effects of Low-level Electrochemical Currents on Bacillus subtilis"

Article Title: Differential Gene Expression to Investigate the Effects of Low-level Electrochemical Currents on Bacillus subtilis

Journal: AMB Express

doi: 10.1186/2191-0855-1-39

Effects of ampicillin on biofilms of B. subtilis 1500 . Biofilms were treated with varying concentrations of ampicillin and 500 μA total DC (83 μA/cm 2 ) concurrently for 15 min at 37°C.
Figure Legend Snippet: Effects of ampicillin on biofilms of B. subtilis 1500 . Biofilms were treated with varying concentrations of ampicillin and 500 μA total DC (83 μA/cm 2 ) concurrently for 15 min at 37°C.

Techniques Used:

Effects of DC and pre-treated media on biofilms of B. subtilis 1500 . Biofilms grown for 2 days on 304L stainless steel electrodes at 37°C were treated with pre-treated LB medium or total applied current for 15 min as indicated. Cell density of the biofilms was calculated from the CFU data.
Figure Legend Snippet: Effects of DC and pre-treated media on biofilms of B. subtilis 1500 . Biofilms grown for 2 days on 304L stainless steel electrodes at 37°C were treated with pre-treated LB medium or total applied current for 15 min as indicated. Cell density of the biofilms was calculated from the CFU data.

Techniques Used:

Effects of electrode material and medium composition on the biofilm cells under DC treatment . Biofilms were grown on graphite electrodes and treated with 500 μA DC current with and without 50 μg/mL ampicillin for 15 min at 37°C as indicated. Modified M56 buffer without chlorine was also tested as the electrolyte solution instead of NaCl buffer or LB medium.
Figure Legend Snippet: Effects of electrode material and medium composition on the biofilm cells under DC treatment . Biofilms were grown on graphite electrodes and treated with 500 μA DC current with and without 50 μg/mL ampicillin for 15 min at 37°C as indicated. Modified M56 buffer without chlorine was also tested as the electrolyte solution instead of NaCl buffer or LB medium.

Techniques Used: Modification

Schematic of the electrochemical cell used in this study . The reference electrode is Ag/AgCl wire inserted in a thin glass tube to prevent contact with the working or counter electrode. Biofilms grown on flat steel or carbon electrodes can be clipped onto the side; the liquid level is about 1 cm below the top of the cuvette when full (3 mL).
Figure Legend Snippet: Schematic of the electrochemical cell used in this study . The reference electrode is Ag/AgCl wire inserted in a thin glass tube to prevent contact with the working or counter electrode. Biofilms grown on flat steel or carbon electrodes can be clipped onto the side; the liquid level is about 1 cm below the top of the cuvette when full (3 mL).

Techniques Used:

28) Product Images from "Anti-caries DNA vaccine-induced secretory immunoglobulin A antibodies inhibit formation of Streptococcus mutans biofilms in vitro"

Article Title: Anti-caries DNA vaccine-induced secretory immunoglobulin A antibodies inhibit formation of Streptococcus mutans biofilms in vitro

Journal: Acta Pharmacologica Sinica

doi: 10.1038/aps.2012.145

The activity of the S-IgA against mature biofilms
Figure Legend Snippet: The activity of the S-IgA against mature biofilms

Techniques Used: Activity Assay

Effect of S-IgA pretreated HA disks on biofilm formation. Acquired pellicle formed by saliva from groups A and B. The pellicle formed by saliva from group A could be observed to have more S-IgA (green) on the HA disks than the pellicle formed by saliva
Figure Legend Snippet: Effect of S-IgA pretreated HA disks on biofilm formation. Acquired pellicle formed by saliva from groups A and B. The pellicle formed by saliva from group A could be observed to have more S-IgA (green) on the HA disks than the pellicle formed by saliva

Techniques Used:

The effect of the S-IgA on the S. mutans biofilm formations on the HA disks
Figure Legend Snippet: The effect of the S-IgA on the S. mutans biofilm formations on the HA disks

Techniques Used:

The effect of the S-IgA pretreated HA disks on biofilm formation
Figure Legend Snippet: The effect of the S-IgA pretreated HA disks on biofilm formation

Techniques Used:

Scanning electron microscopy (SEM) investigation. Surface of hydroxyapatite disks at 2000× (A) and 5000× (B) magnification (HA disk was a standard hexagonal structure). S. mutans 16 h biofilm at 8000× (C) and 20000× (D)
Figure Legend Snippet: Scanning electron microscopy (SEM) investigation. Surface of hydroxyapatite disks at 2000× (A) and 5000× (B) magnification (HA disk was a standard hexagonal structure). S. mutans 16 h biofilm at 8000× (C) and 20000× (D)

Techniques Used: Electron Microscopy

29) Product Images from "Role of the Nuclease of Nontypeable Haemophilus influenzae in Dispersal of Organisms from Biofilms"

Article Title: Role of the Nuclease of Nontypeable Haemophilus influenzae in Dispersal of Organisms from Biofilms

Journal: Infection and Immunity

doi: 10.1128/IAI.02601-14

Evidence that the nuclease is the factor responsible for dispersal of NTHI biofilms. (A) “Comet tails” caused by the release of organisms from NTHI 2019 microcolonies/nascent biofilms as organisms transition from the biofilm to planktonic phase over a 24-h period. (B) Study performed on 2019Δ nuc , which had no evidence of microcolony formation or dispersal of organisms. (C) Partial complementation in cis of 2019Δ nuc because the expression of nuc is unregulated in the complemented strain. Small microcolonies were seen with the comet tail configurations similar to those seen in panel A.
Figure Legend Snippet: Evidence that the nuclease is the factor responsible for dispersal of NTHI biofilms. (A) “Comet tails” caused by the release of organisms from NTHI 2019 microcolonies/nascent biofilms as organisms transition from the biofilm to planktonic phase over a 24-h period. (B) Study performed on 2019Δ nuc , which had no evidence of microcolony formation or dispersal of organisms. (C) Partial complementation in cis of 2019Δ nuc because the expression of nuc is unregulated in the complemented strain. Small microcolonies were seen with the comet tail configurations similar to those seen in panel A.

Techniques Used: Expressing

A 50-image stacked z-series at ×20 magnification of 24-h biofilms grown in continuous flow chambers. The samples were stained with propidium iodide (red) and MAb 6E4 (green) prior to visualization. At 24 h, the NTHI 2019Δ nuc biofilm contains increased amounts of eDNA and large aggregates of organisms (B). This compares to lesser amounts of eDNA and diffuse arrangement of organisms within the biofilm of NTHI 2019 (A) and NTHI 2019Δ nuc :: nuc (C).
Figure Legend Snippet: A 50-image stacked z-series at ×20 magnification of 24-h biofilms grown in continuous flow chambers. The samples were stained with propidium iodide (red) and MAb 6E4 (green) prior to visualization. At 24 h, the NTHI 2019Δ nuc biofilm contains increased amounts of eDNA and large aggregates of organisms (B). This compares to lesser amounts of eDNA and diffuse arrangement of organisms within the biofilm of NTHI 2019 (A) and NTHI 2019Δ nuc :: nuc (C).

Techniques Used: Flow Cytometry, Staining

Cryo sections through 24-h biofilms from NTHI 2019 wild type (A), NTHI 2019Δ nuc (B), and NTHI 2019Δ nuc :: nuc (C). The DNA matrix is stained with DAPI, and the NTHI strains are stained with MAb 6E4. Scale bar, 20 μm. The organisms in the parent strain and in the complemented mutant are clearly dispersed throughout the biofilm, whereas they are clustered in the Δ nuc mutant.
Figure Legend Snippet: Cryo sections through 24-h biofilms from NTHI 2019 wild type (A), NTHI 2019Δ nuc (B), and NTHI 2019Δ nuc :: nuc (C). The DNA matrix is stained with DAPI, and the NTHI strains are stained with MAb 6E4. Scale bar, 20 μm. The organisms in the parent strain and in the complemented mutant are clearly dispersed throughout the biofilm, whereas they are clustered in the Δ nuc mutant.

Techniques Used: Staining, Mutagenesis

Confocal microscopy analysis of 10-μm-thick sections of biofilm formation at day 5 in the chinchilla middle ear after infection with NTHI 2019, NTHI 2019Δ nuc , and NTHI 2019Δ nuc :: nuc . The NTHI are stained with MAb 6E4 (green), and DNA is stained with DRAQ5 (blue). There is an aggregation of organisms staining with MAb 6E4 in NTHI 2019Δ nuc (B) compared to the more diffuse display of organisms in the wild type and the complemented mutant (A and C).
Figure Legend Snippet: Confocal microscopy analysis of 10-μm-thick sections of biofilm formation at day 5 in the chinchilla middle ear after infection with NTHI 2019, NTHI 2019Δ nuc , and NTHI 2019Δ nuc :: nuc . The NTHI are stained with MAb 6E4 (green), and DNA is stained with DRAQ5 (blue). There is an aggregation of organisms staining with MAb 6E4 in NTHI 2019Δ nuc (B) compared to the more diffuse display of organisms in the wild type and the complemented mutant (A and C).

Techniques Used: Confocal Microscopy, Infection, Staining, Mutagenesis

Lateral views and stacked z-series of 48-h biofilms stained with Live/Dead stain (green-red) and DRAQ-5 (blue). (A and C) Biofilm images of NTHI 2019 demonstrating a predominance of live organisms (green), while the images of NTHI 2019Δ nuc biofilm (B and D) demonstrate that the majority of organisms were dead (red) by 48 h.
Figure Legend Snippet: Lateral views and stacked z-series of 48-h biofilms stained with Live/Dead stain (green-red) and DRAQ-5 (blue). (A and C) Biofilm images of NTHI 2019 demonstrating a predominance of live organisms (green), while the images of NTHI 2019Δ nuc biofilm (B and D) demonstrate that the majority of organisms were dead (red) by 48 h.

Techniques Used: Staining

30) Product Images from "Role of de-N-acetylase PgaB from Aggregatibacter actinomycetemcomitans in exopolysaccharide export in biofilm mode of growth"

Article Title: Role of de-N-acetylase PgaB from Aggregatibacter actinomycetemcomitans in exopolysaccharide export in biofilm mode of growth

Journal: Molecular oral microbiology

doi: 10.1111/omi.12188

The role of the catalytic domain of PgaB in phenotypic variation and biofilm integrity. (a) Deletion of catalytic domain of PgaB (ΔNpgaB) results in a phenotype with attenuated rugged edges characteristic of IDH 781 but lacking the internal star
Figure Legend Snippet: The role of the catalytic domain of PgaB in phenotypic variation and biofilm integrity. (a) Deletion of catalytic domain of PgaB (ΔNpgaB) results in a phenotype with attenuated rugged edges characteristic of IDH 781 but lacking the internal star

Techniques Used:

Confocal scanning laser microscopic image of biofilms. (a) Biofilm growth was imaged at seven different locations and averaged for analysis. The scale bar is at 20 μm; (b) Cell viability for IDH 781 and ΔNpgaB strains at different time
Figure Legend Snippet: Confocal scanning laser microscopic image of biofilms. (a) Biofilm growth was imaged at seven different locations and averaged for analysis. The scale bar is at 20 μm; (b) Cell viability for IDH 781 and ΔNpgaB strains at different time

Techniques Used:

Immunofluorescence evaluation of biofilms for the production of PNAG. PNAG was visualized using the human mAb F598 and a secondary anti-human IgG conjugated to Alexa 488 (left panel). Images of the same field viewed by DAPI to stain DNA is shown in the
Figure Legend Snippet: Immunofluorescence evaluation of biofilms for the production of PNAG. PNAG was visualized using the human mAb F598 and a secondary anti-human IgG conjugated to Alexa 488 (left panel). Images of the same field viewed by DAPI to stain DNA is shown in the

Techniques Used: Immunofluorescence, Staining

31) Product Images from " In vitro evaluation of antifungal activity of monolaurin against Candida albicans biofilms"

Article Title: In vitro evaluation of antifungal activity of monolaurin against Candida albicans biofilms

Journal: PeerJ

doi: 10.7717/peerj.2148

Fungal load of biofilms treated with monolaurin and proteolytic enzymes activity level. Fungal load of 1-monolaurin treated biofilms. *p
Figure Legend Snippet: Fungal load of biofilms treated with monolaurin and proteolytic enzymes activity level. Fungal load of 1-monolaurin treated biofilms. *p

Techniques Used: Activity Assay

32) Product Images from "Aggregatibacter actinomycetemcomitans biofilm killing by a targeted ciprofloxacin prodrug"

Article Title: Aggregatibacter actinomycetemcomitans biofilm killing by a targeted ciprofloxacin prodrug

Journal: Biofouling

doi: 10.1080/08927014.2013.823541

Transport of Aa -mAb into biofilm microcolonies. (a) Aa biofilm (3D confocal image, increments on the scale barare 4 μm apart). (b) Transport of Aa -mAb into themicrocolonies of the Aa biofilm based on the green to red ratioof fluorescence (G/R),
Figure Legend Snippet: Transport of Aa -mAb into biofilm microcolonies. (a) Aa biofilm (3D confocal image, increments on the scale barare 4 μm apart). (b) Transport of Aa -mAb into themicrocolonies of the Aa biofilm based on the green to red ratioof fluorescence (G/R),

Techniques Used: Fluorescence

Targeting of Aa biofilm
Figure Legend Snippet: Targeting of Aa biofilm

Techniques Used:

Targeting scheme. (a) Maleimides of the prodrug or fluorescein were linked to abiotinylation reagent via a glutathione coupler withconjugation at the sulfhydryl (arrow). (b) Biotinyated compounds were targetedto the cell wall (CW) of biofilm cells
Figure Legend Snippet: Targeting scheme. (a) Maleimides of the prodrug or fluorescein were linked to abiotinylation reagent via a glutathione coupler withconjugation at the sulfhydryl (arrow). (b) Biotinyated compounds were targetedto the cell wall (CW) of biofilm cells

Techniques Used:

Killing of a biofilm of Aa targeted with prodrug. (a, c) Mediumwithout CMC; (b, d) Medium with CMC. T = targeted, NT =non-targeted. For the non-targeted conditions all the steps in the sequence oftargeting reactions were performed except for the addition
Figure Legend Snippet: Killing of a biofilm of Aa targeted with prodrug. (a, c) Mediumwithout CMC; (b, d) Medium with CMC. T = targeted, NT =non-targeted. For the non-targeted conditions all the steps in the sequence oftargeting reactions were performed except for the addition

Techniques Used: Sequencing

Specific binding of the biotinylated fluorescent analogue and the biotinylatedciprofloxacin prodrug to the biofilm of Aa via thestreptavidin/biotin couple. The fluorescence was normalized to that offluorescently tagged streptavidin added in step iii
Figure Legend Snippet: Specific binding of the biotinylated fluorescent analogue and the biotinylatedciprofloxacin prodrug to the biofilm of Aa via thestreptavidin/biotin couple. The fluorescence was normalized to that offluorescently tagged streptavidin added in step iii

Techniques Used: Binding Assay, Fluorescence

33) Product Images from "Evaluating Streptococcus mutans Strain Dependent Characteristics in a Polymicrobial Biofilm Community"

Article Title: Evaluating Streptococcus mutans Strain Dependent Characteristics in a Polymicrobial Biofilm Community

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.01498

Monospecies cariogenic biofilm biomass is influenced by carbohydrate source. Biomass was quantified spectrophotometrically by reading absorbance at 570 nm in a microtiter plate reader (FluoStar Omega, BMG Labtech). Four replicates were used for each isolate and was carried out on triplicate separate occasions, with the mean of each represented. Data represents mean with significance ∗ p
Figure Legend Snippet: Monospecies cariogenic biofilm biomass is influenced by carbohydrate source. Biomass was quantified spectrophotometrically by reading absorbance at 570 nm in a microtiter plate reader (FluoStar Omega, BMG Labtech). Four replicates were used for each isolate and was carried out on triplicate separate occasions, with the mean of each represented. Data represents mean with significance ∗ p

Techniques Used:

Polymicrobial cariogenic biofilm viability is influenced by carbohydrate source. (A) Polymicrobial caries biofilm model containing standardized L. casei , V. dispar , F. nucleatum , A. naeslundii and S. mutans isolates derived from either caries and caries-free patients grown at was grown in artificial saliva (AS) + galactose or sucrose at 37°C in 5% CO 2 for 5 days, with spent supernatants being replaced with fresh AS every 24 h. Metabolic activity (AlamarBlue) was quantified spectrophotometrically at 490 nm. Significant increase in growth compared to the 24 h culture ( ∗∗ p
Figure Legend Snippet: Polymicrobial cariogenic biofilm viability is influenced by carbohydrate source. (A) Polymicrobial caries biofilm model containing standardized L. casei , V. dispar , F. nucleatum , A. naeslundii and S. mutans isolates derived from either caries and caries-free patients grown at was grown in artificial saliva (AS) + galactose or sucrose at 37°C in 5% CO 2 for 5 days, with spent supernatants being replaced with fresh AS every 24 h. Metabolic activity (AlamarBlue) was quantified spectrophotometrically at 490 nm. Significant increase in growth compared to the 24 h culture ( ∗∗ p

Techniques Used: Derivative Assay, Activity Assay

Polymicrobial cariogenic biofilm biomass is influenced by carbohydrate source. (A) Polymicrobial caries biofilm model containing standardized L. casei , V. dispar , F. nucleatum , A. naeslundii and S. mutans isolates derived from either caries and caries-free patients grown at was grown in artificial saliva (AS) + either galactose or sucrose at 37°C in 5% CO 2 for 5 days, with spent supernatants being replaced with fresh AS every 24 h. Biomass crystal violet staining was quantified spectrophotometrically at 570 nm. Significant increase in growth compared to the 24 h culture ( ∗∗ p
Figure Legend Snippet: Polymicrobial cariogenic biofilm biomass is influenced by carbohydrate source. (A) Polymicrobial caries biofilm model containing standardized L. casei , V. dispar , F. nucleatum , A. naeslundii and S. mutans isolates derived from either caries and caries-free patients grown at was grown in artificial saliva (AS) + either galactose or sucrose at 37°C in 5% CO 2 for 5 days, with spent supernatants being replaced with fresh AS every 24 h. Biomass crystal violet staining was quantified spectrophotometrically at 570 nm. Significant increase in growth compared to the 24 h culture ( ∗∗ p

Techniques Used: Derivative Assay, Staining

34) Product Images from "Role of rpoS in Escherichia coli O157:H7 Strain H32 Biofilm Development and Survival"

Article Title: Role of rpoS in Escherichia coli O157:H7 Strain H32 Biofilm Development and Survival

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.02149-12

Survival of 48-h-old E. coli O157:H7 strain H32- gfp (○) and H32- gfp Δ rpoS :: gm (▼) biofilm cells exposed to sterile distilled water, filter-sterilized lake water, and unfiltered lake water at 22°C. Day 0 represents the time
Figure Legend Snippet: Survival of 48-h-old E. coli O157:H7 strain H32- gfp (○) and H32- gfp Δ rpoS :: gm (▼) biofilm cells exposed to sterile distilled water, filter-sterilized lake water, and unfiltered lake water at 22°C. Day 0 represents the time

Techniques Used:

CSLM images of H32- gfp (A) and H32- gfp Δ rpoS :: gm (B) biofilms in sterile distilled water (ddH 2 O), filter-sterilized lake water (FW), and unfiltered lake water (LW). Day 0 represents the time when E. coli O157:H7 biofilms that were established over
Figure Legend Snippet: CSLM images of H32- gfp (A) and H32- gfp Δ rpoS :: gm (B) biofilms in sterile distilled water (ddH 2 O), filter-sterilized lake water (FW), and unfiltered lake water (LW). Day 0 represents the time when E. coli O157:H7 biofilms that were established over

Techniques Used:

Biofilm development of E. coli O157:H7 strain H32 (●), H32- gfp (○), and H32- gfp Δ rpoS :: gm (▼) in MSMG at 22°C. All three E. coli strains were stained with SYTO 9. Bars indicate standard deviations at a 95% confidence
Figure Legend Snippet: Biofilm development of E. coli O157:H7 strain H32 (●), H32- gfp (○), and H32- gfp Δ rpoS :: gm (▼) in MSMG at 22°C. All three E. coli strains were stained with SYTO 9. Bars indicate standard deviations at a 95% confidence

Techniques Used: Staining

Representative CSLM images of biofilm cells of E. coli O157:H7 strains H32, H32- gfp , and H32- gfp Δ rpoS :: gm (KO) after 24 h of growth in TSB or MSMG. The biofilms were cultivated in TSB at 37°C (left column), MSMG at 37°C (middle
Figure Legend Snippet: Representative CSLM images of biofilm cells of E. coli O157:H7 strains H32, H32- gfp , and H32- gfp Δ rpoS :: gm (KO) after 24 h of growth in TSB or MSMG. The biofilms were cultivated in TSB at 37°C (left column), MSMG at 37°C (middle

Techniques Used:

35) Product Images from "In vitro analysis of flufenamic acid activity against Candida albicans biofilms"

Article Title: In vitro analysis of flufenamic acid activity against Candida albicans biofilms

Journal: International journal of antimicrobial agents

doi: 10.1016/j.ijantimicag.2013.08.018

(a) In vitro effect of flufenamic acid (FFA) against formation (prevention) of Candida albicans biofilms. Planktonic cells were incubated in RPMI 1640 containing FFA at different concentrations (2–1024 mg/L) and biofilms were allowed to form for
Figure Legend Snippet: (a) In vitro effect of flufenamic acid (FFA) against formation (prevention) of Candida albicans biofilms. Planktonic cells were incubated in RPMI 1640 containing FFA at different concentrations (2–1024 mg/L) and biofilms were allowed to form for

Techniques Used: In Vitro, Incubation

36) Product Images from "Effects of CO2 laser irradiation on matrix-rich biofilm development formation–an in vitro study"

Article Title: Effects of CO2 laser irradiation on matrix-rich biofilm development formation–an in vitro study

Journal: PeerJ

doi: 10.7717/peerj.2458

Real time quantitative information about gene expression in S. mutans biofilm after treatments with/without laser irradiation on enamel surface. (A) gtfB (B) gbpB. Values marked by the same letters are not significantly different from each other (p > 0.05). T-test (p > 0.05).
Figure Legend Snippet: Real time quantitative information about gene expression in S. mutans biofilm after treatments with/without laser irradiation on enamel surface. (A) gtfB (B) gbpB. Values marked by the same letters are not significantly different from each other (p > 0.05). T-test (p > 0.05).

Techniques Used: Expressing, Irradiation

Fluorescence Microscopy showing representative images of bacteria in biofilms after 1, 3 and 5 days of biofilm. Multidimensional imaging of live (green) and dead (red) bacteria.
Figure Legend Snippet: Fluorescence Microscopy showing representative images of bacteria in biofilms after 1, 3 and 5 days of biofilm. Multidimensional imaging of live (green) and dead (red) bacteria.

Techniques Used: Fluorescence, Microscopy, Imaging

Morphology and structure of after one, three and five days S. mutans biofilms imaged by FESEM (2500X).
Figure Legend Snippet: Morphology and structure of after one, three and five days S. mutans biofilms imaged by FESEM (2500X).

Techniques Used:

Streptococcus mutans adherence test performed in day 1 of biofilm (expressed in log CFU/mL). Values marked by the distinct letters are significantly different from each other. T-test (p
Figure Legend Snippet: Streptococcus mutans adherence test performed in day 1 of biofilm (expressed in log CFU/mL). Values marked by the distinct letters are significantly different from each other. T-test (p

Techniques Used:

37) Product Images from "The innate immune protein calprotectin promotes Pseudomonas aeruginosa and Staphylococcus aureus interaction"

Article Title: The innate immune protein calprotectin promotes Pseudomonas aeruginosa and Staphylococcus aureus interaction

Journal: Nature Communications

doi: 10.1038/ncomms11951

Differences in metal distribution patterns revealed by LA-ICP IMS correlate with differential protein localization within the biofilm. ( a ) LA-ICP IMS analysis of a biofilm section containing a defined ‘central channel' and ‘nutrient-deplete edge'. Scale bar, 2 mm. ( b ) ICP-MS quantification of metal levels within distinct portions of the biofilm. Error bars represent s.d. of data derived from triplicate biofilms. ‘*' designates P
Figure Legend Snippet: Differences in metal distribution patterns revealed by LA-ICP IMS correlate with differential protein localization within the biofilm. ( a ) LA-ICP IMS analysis of a biofilm section containing a defined ‘central channel' and ‘nutrient-deplete edge'. Scale bar, 2 mm. ( b ) ICP-MS quantification of metal levels within distinct portions of the biofilm. Error bars represent s.d. of data derived from triplicate biofilms. ‘*' designates P

Techniques Used: Mass Spectrometry, Derivative Assay

MALDI IMS signals identified using bottom-up proteomics. m/z values and associated protein designations are included adjacent to MALDI IMS images of the biofilm section proximal to the nutrient pore highlighted in Fig. 1 . These identifications derive from signals that were reproducibly detected during the analysis of 15 replicate DFR biofilms. Scale bar, 3 mm.
Figure Legend Snippet: MALDI IMS signals identified using bottom-up proteomics. m/z values and associated protein designations are included adjacent to MALDI IMS images of the biofilm section proximal to the nutrient pore highlighted in Fig. 1 . These identifications derive from signals that were reproducibly detected during the analysis of 15 replicate DFR biofilms. Scale bar, 3 mm.

Techniques Used:

Bottom-up proteomics reveals repression of anti-staphylococcal biosynthetic proteins in the edge of the P. aeruginosa biofilm. HcnB, PhzS and PqsB are components of various anti-staphylococcal biosynthetic pathways that were found in lower abundance in the biofilm edge. While expression of the corresponding genes has been shown to be both Fe and quorum sensing (QS)-regulated, the proteins' distribution profiles did not correlate with those of known Fe-responsive proteins such as FpvA and SdhA or proteins encoded by known QS-responsive genes such as LasA and RmlC. Error bars are s.e.m. derived from biological triplicate samples processed in four technical replicates per biofilm. ‘*' denotes P
Figure Legend Snippet: Bottom-up proteomics reveals repression of anti-staphylococcal biosynthetic proteins in the edge of the P. aeruginosa biofilm. HcnB, PhzS and PqsB are components of various anti-staphylococcal biosynthetic pathways that were found in lower abundance in the biofilm edge. While expression of the corresponding genes has been shown to be both Fe and quorum sensing (QS)-regulated, the proteins' distribution profiles did not correlate with those of known Fe-responsive proteins such as FpvA and SdhA or proteins encoded by known QS-responsive genes such as LasA and RmlC. Error bars are s.e.m. derived from biological triplicate samples processed in four technical replicates per biofilm. ‘*' denotes P

Techniques Used: Expressing, Derivative Assay

Heterogeneous structure of a Pseudomonas aeruginosa biofilm grown in a DFR. ( a ) Representative image of a P. aeruginosa biofilm grown in a drip flow reactor (DFR). The flow of nutrients is depicted by the topmost arrows ranging in colour from blue to brown. Blue boxes denote the approximate regions from which 12-μm-thick sections were obtained for MALDI IMS analysis. ( b , c ) MALDI IMS signals with differential biofilm localization found primarily in portions of the biomass presumed to be nutrient-replete. ( d , e ) MALDI IMS signals primarily localized to portions of the biomass predicted to be nutrient-deplete. ( f ) Overlay of signals shown in b – e highlighting sublocalization of signals within the predicted nutrient-replete and nutrient-deplete niches with zoomed insets highlighted. Scale bar, 3 mm (inset scale bars, 1 mm).
Figure Legend Snippet: Heterogeneous structure of a Pseudomonas aeruginosa biofilm grown in a DFR. ( a ) Representative image of a P. aeruginosa biofilm grown in a drip flow reactor (DFR). The flow of nutrients is depicted by the topmost arrows ranging in colour from blue to brown. Blue boxes denote the approximate regions from which 12-μm-thick sections were obtained for MALDI IMS analysis. ( b , c ) MALDI IMS signals with differential biofilm localization found primarily in portions of the biomass presumed to be nutrient-replete. ( d , e ) MALDI IMS signals primarily localized to portions of the biomass predicted to be nutrient-deplete. ( f ) Overlay of signals shown in b – e highlighting sublocalization of signals within the predicted nutrient-replete and nutrient-deplete niches with zoomed insets highlighted. Scale bar, 3 mm (inset scale bars, 1 mm).

Techniques Used: Flow Cytometry

38) Product Images from "Mechanism of Fluconazole Resistance in Candida albicans Biofilms: Phase-Specific Role of Efflux Pumps and Membrane Sterols "

Article Title: Mechanism of Fluconazole Resistance in Candida albicans Biofilms: Phase-Specific Role of Efflux Pumps and Membrane Sterols

Journal: Infection and Immunity

doi: 10.1128/IAI.71.8.4333-4340.2003

Expression of CDR and MDR1 genes in (P) planktonic (P) and biofilm (B) forms of C. albicans . Total RNA was isolated from biofilms and planktonic cells grown for 6, 12, and 48 h. Sixty micrograms of total RNA was analyzed by Northern blotting using a CDR - or MDR1 -specific probe as described in Materials and Methods. Top, CDR transcript; middle, MDR1 transcript; bottom, 25S rRNA (loading control). Results are representative of three separate experiments.
Figure Legend Snippet: Expression of CDR and MDR1 genes in (P) planktonic (P) and biofilm (B) forms of C. albicans . Total RNA was isolated from biofilms and planktonic cells grown for 6, 12, and 48 h. Sixty micrograms of total RNA was analyzed by Northern blotting using a CDR - or MDR1 -specific probe as described in Materials and Methods. Top, CDR transcript; middle, MDR1 transcript; bottom, 25S rRNA (loading control). Results are representative of three separate experiments.

Techniques Used: Expressing, Isolation, Northern Blot

Percent growth inhibition of C. albicans biofilms exposed to high concentration of fluconazole. Percentages of inhibition for biofilms grown to the early (6 h), intermediate (12 h), or late (48 h) phase of development and exposed to 256 μg of fluconazole/ml were determined. Strains used were: CAF2-1 (wild type), DSY448 (Δ cdr1 ), DSY465 (Δ mdr1 ), DSY654 (Δ cdr1 Δ cdr2 ), and DSY1050 (Δ cdr1 Δ cdr2 Δ mdr1 ). For each strain, drug susceptibility decreased from the early to late phase of biofilm development. Additionally, deletion of two and three efflux pumps led to progressively decreasing susceptibility to fluconazole. Metabolic activity was normalized to the control without fluconazole, which was taken as 100%. Data (means ± standard deviations) are representative of three separate experiments. ∗, the wild-type (CAF2-1) strain showed 0% inhibition at all the time points.
Figure Legend Snippet: Percent growth inhibition of C. albicans biofilms exposed to high concentration of fluconazole. Percentages of inhibition for biofilms grown to the early (6 h), intermediate (12 h), or late (48 h) phase of development and exposed to 256 μg of fluconazole/ml were determined. Strains used were: CAF2-1 (wild type), DSY448 (Δ cdr1 ), DSY465 (Δ mdr1 ), DSY654 (Δ cdr1 Δ cdr2 ), and DSY1050 (Δ cdr1 Δ cdr2 Δ mdr1 ). For each strain, drug susceptibility decreased from the early to late phase of biofilm development. Additionally, deletion of two and three efflux pumps led to progressively decreasing susceptibility to fluconazole. Metabolic activity was normalized to the control without fluconazole, which was taken as 100%. Data (means ± standard deviations) are representative of three separate experiments. ∗, the wild-type (CAF2-1) strain showed 0% inhibition at all the time points.

Techniques Used: Inhibition, Concentration Assay, Activity Assay

Rh123 accumulation by early-, intermediate-, and mature-phase biofilms and planktonic cells of C. albicans. Data were analyzed by two-way analysis of variance, and a value of P
Figure Legend Snippet: Rh123 accumulation by early-, intermediate-, and mature-phase biofilms and planktonic cells of C. albicans. Data were analyzed by two-way analysis of variance, and a value of P

Techniques Used:

Variations in sterol profiles of C. albicans biofilms at different developmental phases. Sterol patterns for biofilms grown to the early (A), intermediate (B), or mature (C) phase were determined by GLC. (D) Percentages of sterols identified in C. albicans biofilms and planktonic cells (chromatograph not shown), determined from the corresponding peak areas and retention times relative to ergosterol. Peaks 1 to 7 (A to C) represent sterols described in panel D. SD, standard deviation.
Figure Legend Snippet: Variations in sterol profiles of C. albicans biofilms at different developmental phases. Sterol patterns for biofilms grown to the early (A), intermediate (B), or mature (C) phase were determined by GLC. (D) Percentages of sterols identified in C. albicans biofilms and planktonic cells (chromatograph not shown), determined from the corresponding peak areas and retention times relative to ergosterol. Peaks 1 to 7 (A to C) represent sterols described in panel D. SD, standard deviation.

Techniques Used: Gas Chromatography, Standard Deviation

39) Product Images from "Differential Gene Expression to Investigate the Effects of Low-level Electrochemical Currents on Bacillus subtilis"

Article Title: Differential Gene Expression to Investigate the Effects of Low-level Electrochemical Currents on Bacillus subtilis

Journal: AMB Express

doi: 10.1186/2191-0855-1-39

Effects of ampicillin on biofilms of B. subtilis 1500 . Biofilms were treated with varying concentrations of ampicillin and 500 μA total DC (83 μA/cm 2 ) concurrently for 15 min at 37°C.
Figure Legend Snippet: Effects of ampicillin on biofilms of B. subtilis 1500 . Biofilms were treated with varying concentrations of ampicillin and 500 μA total DC (83 μA/cm 2 ) concurrently for 15 min at 37°C.

Techniques Used:

Effects of DC and pre-treated media on biofilms of B. subtilis 1500 . Biofilms grown for 2 days on 304L stainless steel electrodes at 37°C were treated with pre-treated LB medium or total applied current for 15 min as indicated. Cell density of the biofilms was calculated from the CFU data.
Figure Legend Snippet: Effects of DC and pre-treated media on biofilms of B. subtilis 1500 . Biofilms grown for 2 days on 304L stainless steel electrodes at 37°C were treated with pre-treated LB medium or total applied current for 15 min as indicated. Cell density of the biofilms was calculated from the CFU data.

Techniques Used:

Effects of electrode material and medium composition on the biofilm cells under DC treatment . Biofilms were grown on graphite electrodes and treated with 500 μA DC current with and without 50 μg/mL ampicillin for 15 min at 37°C as indicated. Modified M56 buffer without chlorine was also tested as the electrolyte solution instead of NaCl buffer or LB medium.
Figure Legend Snippet: Effects of electrode material and medium composition on the biofilm cells under DC treatment . Biofilms were grown on graphite electrodes and treated with 500 μA DC current with and without 50 μg/mL ampicillin for 15 min at 37°C as indicated. Modified M56 buffer without chlorine was also tested as the electrolyte solution instead of NaCl buffer or LB medium.

Techniques Used: Modification

Schematic of the electrochemical cell used in this study . The reference electrode is Ag/AgCl wire inserted in a thin glass tube to prevent contact with the working or counter electrode. Biofilms grown on flat steel or carbon electrodes can be clipped onto the side; the liquid level is about 1 cm below the top of the cuvette when full (3 mL).
Figure Legend Snippet: Schematic of the electrochemical cell used in this study . The reference electrode is Ag/AgCl wire inserted in a thin glass tube to prevent contact with the working or counter electrode. Biofilms grown on flat steel or carbon electrodes can be clipped onto the side; the liquid level is about 1 cm below the top of the cuvette when full (3 mL).

Techniques Used:

40) Product Images from "Effect of Twice-Daily Blue Light Treatment on Matrix-Rich Biofilm Development"

Article Title: Effect of Twice-Daily Blue Light Treatment on Matrix-Rich Biofilm Development

Journal: PLoS ONE

doi: 10.1371/journal.pone.0131941

CFU count and dry weight in S . mutans biofilm. CFU count and dry weight in the S . mutans biofilm after the twice-daily blue light treatment compared with twice daily treatment with 0.12% Chlorhexidine (positive control) and twice-daily treatment with 0.89% NaCl (negative control). Data represent the mean values and error bars represent standard deviations. Values marked by the different letters are significantly different from each other (p
Figure Legend Snippet: CFU count and dry weight in S . mutans biofilm. CFU count and dry weight in the S . mutans biofilm after the twice-daily blue light treatment compared with twice daily treatment with 0.12% Chlorhexidine (positive control) and twice-daily treatment with 0.89% NaCl (negative control). Data represent the mean values and error bars represent standard deviations. Values marked by the different letters are significantly different from each other (p

Techniques Used: Positive Control, Negative Control

Morphology and structure of S . mutans biofilms imaged by VPSEM. VPSEM images are showing the morphology and structure of S . mutans biofilm after the twice-daily blue light treatment compared with positive and negative controls [Field width of 1.0 mm]. A = Biofilm after the twice daily blue light treatment; B = Biofilm after treatment with 0.12% chlorhexidine (positive control); C = Biofilm after treatment with 0.89% NaCl (negative control).
Figure Legend Snippet: Morphology and structure of S . mutans biofilms imaged by VPSEM. VPSEM images are showing the morphology and structure of S . mutans biofilm after the twice-daily blue light treatment compared with positive and negative controls [Field width of 1.0 mm]. A = Biofilm after the twice daily blue light treatment; B = Biofilm after treatment with 0.12% chlorhexidine (positive control); C = Biofilm after treatment with 0.89% NaCl (negative control).

Techniques Used: Positive Control, Negative Control

Viability and spatial arrangement in the S . mutans biofilms showed by confocal laser scanning microscopy. Five (5) day-old biofilms were stained with BacLight LIVE/DEAD and processed for CSLM. (A) Biofilm after the twice-daily blue light treatment during 5 days. (B) Biofilm after twice-daily treatment with 0.12% Chlorhexidine (positive control). (C) Biofilm after twice-daily treatment with 0.89% NaCl (negative control). (1) Red cells are considered dead and green cells are alive. (2) Overlap of live and dead stacks. (3) Orthogonal view of the overlap of live and dead stacks; dots line represents the interface biofilm/ HA substrate. (4) Representative three-dimensional images of the structural organization of the biofilms: rendered images of the outer layers of biofilms. The total bacterial biomasses calculated by COMSTAT are shown.
Figure Legend Snippet: Viability and spatial arrangement in the S . mutans biofilms showed by confocal laser scanning microscopy. Five (5) day-old biofilms were stained with BacLight LIVE/DEAD and processed for CSLM. (A) Biofilm after the twice-daily blue light treatment during 5 days. (B) Biofilm after twice-daily treatment with 0.12% Chlorhexidine (positive control). (C) Biofilm after twice-daily treatment with 0.89% NaCl (negative control). (1) Red cells are considered dead and green cells are alive. (2) Overlap of live and dead stacks. (3) Orthogonal view of the overlap of live and dead stacks; dots line represents the interface biofilm/ HA substrate. (4) Representative three-dimensional images of the structural organization of the biofilms: rendered images of the outer layers of biofilms. The total bacterial biomasses calculated by COMSTAT are shown.

Techniques Used: Confocal Laser Scanning Microscopy, Staining, Positive Control, Negative Control

The content of EPS-soluble, EPS-insoluble and IPS in S . mutans biofilm. The content of soluble and insoluble extracellular polysaccharides (EPS-soluble and EPS-insoluble, respectively), and of intracellular polysaccharides (IPS) in S . mutans biofilm (expressed in μg/mg of biofilm) after the twice-daily blue light treatment compared with twice daily treatment with 0.12% Chlorhexidine (positive control) and twice-daily treatment with 0.89% NaCl (negative control). Data represent the mean values and error bars represent standard deviations. Values marked by the different letters are significantly different from each other (p
Figure Legend Snippet: The content of EPS-soluble, EPS-insoluble and IPS in S . mutans biofilm. The content of soluble and insoluble extracellular polysaccharides (EPS-soluble and EPS-insoluble, respectively), and of intracellular polysaccharides (IPS) in S . mutans biofilm (expressed in μg/mg of biofilm) after the twice-daily blue light treatment compared with twice daily treatment with 0.12% Chlorhexidine (positive control) and twice-daily treatment with 0.89% NaCl (negative control). Data represent the mean values and error bars represent standard deviations. Values marked by the different letters are significantly different from each other (p

Techniques Used: Positive Control, Negative Control

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Article Title: Nontypeable Haemophilus influenzae Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate In Vivo, Which Can Be Used as a Target for Bactericidal Antibody
Article Snippet: .. To preserve the architecture of biofilms that had formed in vivo , we embedded the biofilm sample in an OCT compound (Fisher Scientific, Pittsburgh, PA) as previously described ( ). .. Serial sections (4-µm thickness) were cut on a Leica CM3050S cryotome (Leica Microsystems, Inc., Bannockburn, IL).

Generated:

Article Title: The Cyclic AMP-Dependent Catabolite Repression System of Serratia marcescens Mediates Biofilm Formation through Regulation of Type 1 Fimbriae
Article Snippet: .. Biofilms on glass were generated using 20- by 150-mm borosilicate glass test tubes (14-961-33; Fisher Scientific) with 5 ml of LB incubated overnight at 30°C on a TC-7 tissue culture roller at full speed, 56 rpm (New Brunswick Instruments). .. These biofilms were stained while rotating, using 6 ml of 0.1% crystal violet, and solubilized using 6 ml of 30% glacial acetic acid, and absorbance for 150-μl aliquots was determined at 590 nm with a Synergy 2 plate reader (Biotek).

Incubation:

Article Title: The Cyclic AMP-Dependent Catabolite Repression System of Serratia marcescens Mediates Biofilm Formation through Regulation of Type 1 Fimbriae
Article Snippet: .. Biofilms on glass were generated using 20- by 150-mm borosilicate glass test tubes (14-961-33; Fisher Scientific) with 5 ml of LB incubated overnight at 30°C on a TC-7 tissue culture roller at full speed, 56 rpm (New Brunswick Instruments). .. These biofilms were stained while rotating, using 6 ml of 0.1% crystal violet, and solubilized using 6 ml of 30% glacial acetic acid, and absorbance for 150-μl aliquots was determined at 590 nm with a Synergy 2 plate reader (Biotek).

Article Title: Rhesus θ-defensin-1 (RTD-1) exhibits in vitro and in vivo activity against cystic fibrosis strains of Pseudomonas aeruginosa
Article Snippet: .. After 48 h of growth on M63 agarose alone, biofilms were transferred to the RTD-1-containing plates and incubated for 24 h, then transferred to a fresh RTD-1-containing plate and incubated for an additional 24 h. Biofilms were aseptically removed, placed in 10 mL of sterile PBS and subjected to sonication for 10 min in a Solid State Ultrasonic FS-9 sonicator (Fisher Scientific, Pittsburg, PA, USA) at 4°C for 10 min with an additional 2 min of vortexing, until complete removal of biofilms from the filters was achieved. ..

Sonication:

Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions
Article Snippet: .. After growth, biofilms were gently washed with PBS before being treated with 40 mg/L of miconazole for a further 24 h. Following treatment, biofilms were washed before being sonicated for 5 min in 1 mL of PBS at 35 kHz in an ultrasonic water-bath (Fisher Scientific, Leicestershire, UK) to remove biofilm cells, and the viability of the biofilms after treatment was determined using species-specific live/dead PCR, as described above. ..

Article Title: Propolis Is an Efficient Fungicide and Inhibitor of Biofilm Production by Vaginal Candida albicans
Article Snippet: .. Briefly, the biofilm samples were scraped from the 24-well plates, resuspended with ultra-pure water, and sonicated (Sonic Dismembrator Ultrasonic Processor, Fisher Scientific) for 45 s at 30 W, and then the suspension was vortexed for 2 min. ..

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Article Snippet: .. After 48 h of growth on M63 agarose alone, biofilms were transferred to the RTD-1-containing plates and incubated for 24 h, then transferred to a fresh RTD-1-containing plate and incubated for an additional 24 h. Biofilms were aseptically removed, placed in 10 mL of sterile PBS and subjected to sonication for 10 min in a Solid State Ultrasonic FS-9 sonicator (Fisher Scientific, Pittsburg, PA, USA) at 4°C for 10 min with an additional 2 min of vortexing, until complete removal of biofilms from the filters was achieved. ..

Polymerase Chain Reaction:

Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions
Article Snippet: .. After growth, biofilms were gently washed with PBS before being treated with 40 mg/L of miconazole for a further 24 h. Following treatment, biofilms were washed before being sonicated for 5 min in 1 mL of PBS at 35 kHz in an ultrasonic water-bath (Fisher Scientific, Leicestershire, UK) to remove biofilm cells, and the viability of the biofilms after treatment was determined using species-specific live/dead PCR, as described above. ..

Staining:

Article Title: Searching for new strategies against biofilm infections: Colistin-AMP combinations against Pseudomonas aeruginosa and Staphylococcus aureus single- and double-species biofilms
Article Snippet: .. The plates were emptied by plate inversion, air dried and the stain bound to the adherent biofilm was re-suspended with 200 μL of 33% (V/V) glacial acetic acid (Fischer Scientific). ..

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    Fisher Scientific biofilm
    Scanning electron microscopy images of the effect of the PES on Candida albicans <t>biofilm</t> on a polystyrene surface for three samples: A2 isolate ((a), (b)), 31MC isolate ((c), (d)), and ATCC90028 reference strain ((e), (f)). Biofilms without the PES ((a), (c), and (e)) and biofilms with the PES (273.43 μ g/mL of total phenol content in gallic acid) after 24 h ((b), (d), and (f)) at 350x magnification.
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    Fisher Scientific static biofilms
    sarA and extracellular proteases significantly affect <t>biofilm</t> formation in a CNS catheter infection model. Infected catheters were removed, rinsed and sonicated for quantification of viable bacteria associated with wild type USA300 LAC ( WT catheter ), USA300 LACΔ sarA ( SarA-deficient catheter ), extracellular protease deficient USA300 LAC ( Protease-deficient catheter ), or SarA and extracellular protease deficient ( SarA-Protease-deficient catheter ) S. aureus . At all time points, bacterial burdens in brain parenchyma were 3–5 log lower than the corresponding catheter cultures (See Figure S4 ). a = p
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    Fisher Scientific biofilm sampling
    Correlation of V. vermiformis with ( A ) Legionella spp. and ( B ) L. pneumophila in distal tap <t>biofilms.</t>
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    Image Search Results


    Scanning electron microscopy images of the effect of the PES on Candida albicans biofilm on a polystyrene surface for three samples: A2 isolate ((a), (b)), 31MC isolate ((c), (d)), and ATCC90028 reference strain ((e), (f)). Biofilms without the PES ((a), (c), and (e)) and biofilms with the PES (273.43 μ g/mL of total phenol content in gallic acid) after 24 h ((b), (d), and (f)) at 350x magnification.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Propolis Is an Efficient Fungicide and Inhibitor of Biofilm Production by Vaginal Candida albicans

    doi: 10.1155/2015/287693

    Figure Lengend Snippet: Scanning electron microscopy images of the effect of the PES on Candida albicans biofilm on a polystyrene surface for three samples: A2 isolate ((a), (b)), 31MC isolate ((c), (d)), and ATCC90028 reference strain ((e), (f)). Biofilms without the PES ((a), (c), and (e)) and biofilms with the PES (273.43 μ g/mL of total phenol content in gallic acid) after 24 h ((b), (d), and (f)) at 350x magnification.

    Article Snippet: Briefly, the biofilm samples were scraped from the 24-well plates, resuspended with ultra-pure water, and sonicated (Sonic Dismembrator Ultrasonic Processor, Fisher Scientific) for 45 s at 30 W, and then the suspension was vortexed for 2 min.

    Techniques: Electron Microscopy

    Antibiofilm effects of zinc pyrithione and silver sulfadiazine individually and in combination against S. aureus , A. baumannii , and P. aeruginosa . The log decrease in the number of biofilm-associated cells following treatment with 2-fold-increasing concentrations (0 to 128 µg·ml −1 ) of silver sulfadiazine (gray bars), zinc pyrithione (black bars), or silver sulfadiazine-zinc pyrithione combination (white bars) in comparison to mock-treated (DMSO) biofilms for S. aureus (A), A. baumannii (B), and P. aeruginosa (C). Error bars represent standard deviations. Significant differences between the results seen with silver sulfadiazine and silver sulfadiazine-zinc pyrithione are indicated. *, P ≤ 0.05; **, P ≤ 0.01 (Student’s t test).

    Journal: mSphere

    Article Title: Zinc Pyrithione Improves the Antibacterial Activity of Silver Sulfadiazine Ointment

    doi: 10.1128/mSphere.00194-16

    Figure Lengend Snippet: Antibiofilm effects of zinc pyrithione and silver sulfadiazine individually and in combination against S. aureus , A. baumannii , and P. aeruginosa . The log decrease in the number of biofilm-associated cells following treatment with 2-fold-increasing concentrations (0 to 128 µg·ml −1 ) of silver sulfadiazine (gray bars), zinc pyrithione (black bars), or silver sulfadiazine-zinc pyrithione combination (white bars) in comparison to mock-treated (DMSO) biofilms for S. aureus (A), A. baumannii (B), and P. aeruginosa (C). Error bars represent standard deviations. Significant differences between the results seen with silver sulfadiazine and silver sulfadiazine-zinc pyrithione are indicated. *, P ≤ 0.05; **, P ≤ 0.01 (Student’s t test).

    Article Snippet: Plates were incubated for 24 h, at which point the supernatant was removed, the biofilm was resuspended in 0.8% NaCl, and the biofilm-associated cells were enumerated by plating on MH agar (Fisher Scientific).

    Techniques:

    sarA and extracellular proteases significantly affect biofilm formation in a CNS catheter infection model. Infected catheters were removed, rinsed and sonicated for quantification of viable bacteria associated with wild type USA300 LAC ( WT catheter ), USA300 LACΔ sarA ( SarA-deficient catheter ), extracellular protease deficient USA300 LAC ( Protease-deficient catheter ), or SarA and extracellular protease deficient ( SarA-Protease-deficient catheter ) S. aureus . At all time points, bacterial burdens in brain parenchyma were 3–5 log lower than the corresponding catheter cultures (See Figure S4 ). a = p

    Journal: PLoS ONE

    Article Title: Staphylococcus aureus sarA Regulates Inflammation and Colonization during Central Nervous System Biofilm Formation

    doi: 10.1371/journal.pone.0084089

    Figure Lengend Snippet: sarA and extracellular proteases significantly affect biofilm formation in a CNS catheter infection model. Infected catheters were removed, rinsed and sonicated for quantification of viable bacteria associated with wild type USA300 LAC ( WT catheter ), USA300 LACΔ sarA ( SarA-deficient catheter ), extracellular protease deficient USA300 LAC ( Protease-deficient catheter ), or SarA and extracellular protease deficient ( SarA-Protease-deficient catheter ) S. aureus . At all time points, bacterial burdens in brain parenchyma were 3–5 log lower than the corresponding catheter cultures (See Figure S4 ). a = p

    Article Snippet: For confocal microscopy analysis, static biofilms were grown on sterile glass chamber slides (Fisher Scientific, Houston TX) treated with 20% human plasma (generous gift of Dr. Steve Carson, UNMC) in sterile carbonate-bicarbonate buffer overnight .

    Techniques: Infection, Sonication

    CNS catheter-associated biofilm infection is associated with attenuated inflammation compared to parenchymal abscesses. The tissues encompassing the brain abscess and the tissues surrounding the infected catheters were homogenized and the resulting supernatants analyzed for levels of the pro-inflammatory mediators CXCL1 (A, B) and IL-17 (C, D). Raw cytokine/chemokine levels are reported (A, C) as well as correction for disparate bacterial burdens (B, D) * = p

    Journal: PLoS ONE

    Article Title: Staphylococcus aureus sarA Regulates Inflammation and Colonization during Central Nervous System Biofilm Formation

    doi: 10.1371/journal.pone.0084089

    Figure Lengend Snippet: CNS catheter-associated biofilm infection is associated with attenuated inflammation compared to parenchymal abscesses. The tissues encompassing the brain abscess and the tissues surrounding the infected catheters were homogenized and the resulting supernatants analyzed for levels of the pro-inflammatory mediators CXCL1 (A, B) and IL-17 (C, D). Raw cytokine/chemokine levels are reported (A, C) as well as correction for disparate bacterial burdens (B, D) * = p

    Article Snippet: For confocal microscopy analysis, static biofilms were grown on sterile glass chamber slides (Fisher Scientific, Houston TX) treated with 20% human plasma (generous gift of Dr. Steve Carson, UNMC) in sterile carbonate-bicarbonate buffer overnight .

    Techniques: Infection

    sarA is critical for catheter-associated biofilm formation in a CNS catheter infection model. Infected catheters were removed, rinsed and sonicated for quantification of viable bacteria associated with wild type ACH1719 ( WT ) or ACH1719Δ sarA ( Δ sarA ) S. aureus . At all time points, bacterial burdens in brain parenchyma were lower than the corresponding catheter cultures (See Figure S2 ). * = p

    Journal: PLoS ONE

    Article Title: Staphylococcus aureus sarA Regulates Inflammation and Colonization during Central Nervous System Biofilm Formation

    doi: 10.1371/journal.pone.0084089

    Figure Lengend Snippet: sarA is critical for catheter-associated biofilm formation in a CNS catheter infection model. Infected catheters were removed, rinsed and sonicated for quantification of viable bacteria associated with wild type ACH1719 ( WT ) or ACH1719Δ sarA ( Δ sarA ) S. aureus . At all time points, bacterial burdens in brain parenchyma were lower than the corresponding catheter cultures (See Figure S2 ). * = p

    Article Snippet: For confocal microscopy analysis, static biofilms were grown on sterile glass chamber slides (Fisher Scientific, Houston TX) treated with 20% human plasma (generous gift of Dr. Steve Carson, UNMC) in sterile carbonate-bicarbonate buffer overnight .

    Techniques: Infection, Sonication

    MSSA CNS catheter-associated biofilm infection persists longer than a parenchymal brain abscess. The infected tissue was removed, homogenized and cultured to enumerate bacterial burdens in the tissue containing the brain abscess or surrounding the infected catheter. Catheters were also removed, rinsed and sonicated for quantification of viable bacteria. * = p

    Journal: PLoS ONE

    Article Title: Staphylococcus aureus sarA Regulates Inflammation and Colonization during Central Nervous System Biofilm Formation

    doi: 10.1371/journal.pone.0084089

    Figure Lengend Snippet: MSSA CNS catheter-associated biofilm infection persists longer than a parenchymal brain abscess. The infected tissue was removed, homogenized and cultured to enumerate bacterial burdens in the tissue containing the brain abscess or surrounding the infected catheter. Catheters were also removed, rinsed and sonicated for quantification of viable bacteria. * = p

    Article Snippet: For confocal microscopy analysis, static biofilms were grown on sterile glass chamber slides (Fisher Scientific, Houston TX) treated with 20% human plasma (generous gift of Dr. Steve Carson, UNMC) in sterile carbonate-bicarbonate buffer overnight .

    Techniques: Infection, Cell Culture, Sonication

    Correlation of V. vermiformis with ( A ) Legionella spp. and ( B ) L. pneumophila in distal tap biofilms.

    Journal: Pathogens

    Article Title: Convective Mixing in Distal Pipes Exacerbates Legionella pneumophila Growth in Hot Water Plumbing

    doi: 10.3390/pathogens5010029

    Figure Lengend Snippet: Correlation of V. vermiformis with ( A ) Legionella spp. and ( B ) L. pneumophila in distal tap biofilms.

    Article Snippet: For biofilm sampling, 65 cm2 of influent, recirculating line, and ends of the distal tap pipes accessible by threaded union connections were swabbed using sterile cotton-tip applicators (Fisherbrand, Fisher Scientific, UK).

    Techniques: