Structured Review

Avantor biofilms
Confocal visual effect of heat on P. aeruginosa <t>biofilms.</t> P. aeruginosa biofilms grown in a drip flow reactor were heat shocked and visualized with Syto9 and propidium iodide dyes under a CLSM. (A) The control heat shock at 37 °C for 2 min had more green fluorescence (live cells dyed with Syto9) than (B) the biofilm heat shocked at 80 °C for 1 min which had more red fluorescence (dead cells dyed with propidium iodide).
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1) Product Images from "Synergistic effects of heat and antibiotics on Pseudomonas aeruginosa biofilms"

Article Title: Synergistic effects of heat and antibiotics on Pseudomonas aeruginosa biofilms

Journal: Biofouling

doi: 10.1080/08927014.2017.1381688

Confocal visual effect of heat on P. aeruginosa biofilms. P. aeruginosa biofilms grown in a drip flow reactor were heat shocked and visualized with Syto9 and propidium iodide dyes under a CLSM. (A) The control heat shock at 37 °C for 2 min had more green fluorescence (live cells dyed with Syto9) than (B) the biofilm heat shocked at 80 °C for 1 min which had more red fluorescence (dead cells dyed with propidium iodide).
Figure Legend Snippet: Confocal visual effect of heat on P. aeruginosa biofilms. P. aeruginosa biofilms grown in a drip flow reactor were heat shocked and visualized with Syto9 and propidium iodide dyes under a CLSM. (A) The control heat shock at 37 °C for 2 min had more green fluorescence (live cells dyed with Syto9) than (B) the biofilm heat shocked at 80 °C for 1 min which had more red fluorescence (dead cells dyed with propidium iodide).

Techniques Used: Flow Cytometry, Confocal Laser Scanning Microscopy, Fluorescence

Effect of heat and tobramycin on the biofilm. Biofilms were incubated at 37 °C with the indicated tobramycin concentrations for 24 h before enumeration. 4 h into this incubation the biofilms were heat shocked for the indicated time and temperature (in some cases at 37 °C as controls). Error bars indicate the SD (n = 12). Asterisks indicate trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p
Figure Legend Snippet: Effect of heat and tobramycin on the biofilm. Biofilms were incubated at 37 °C with the indicated tobramycin concentrations for 24 h before enumeration. 4 h into this incubation the biofilms were heat shocked for the indicated time and temperature (in some cases at 37 °C as controls). Error bars indicate the SD (n = 12). Asterisks indicate trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p

Techniques Used: Incubation, Concentration Assay

Biofilm growth and combined heat shock and antibiotic treatment. Stock P. aeruginosa PAO1 cells were prepared to produce biofilms which were grown in an MBEC™ assay on an orbital shaker table. The biofilms were then transferred via the peg lid to a rinse plate to rinse off any loosely adhered bacteria then moved over to the challenge plate containing antibiotics and controls in different wells. After 4 h of the initial challenge plate the biofilms were heat shocked in a heated water bath and then swiftly transferred to a new challenge plate for the remainder of the total 24 h antibiotic exposure. The biofilms were rinsed once again and then placed in a recovery plate for sonication, dilution, and enumeration.
Figure Legend Snippet: Biofilm growth and combined heat shock and antibiotic treatment. Stock P. aeruginosa PAO1 cells were prepared to produce biofilms which were grown in an MBEC™ assay on an orbital shaker table. The biofilms were then transferred via the peg lid to a rinse plate to rinse off any loosely adhered bacteria then moved over to the challenge plate containing antibiotics and controls in different wells. After 4 h of the initial challenge plate the biofilms were heat shocked in a heated water bath and then swiftly transferred to a new challenge plate for the remainder of the total 24 h antibiotic exposure. The biofilms were rinsed once again and then placed in a recovery plate for sonication, dilution, and enumeration.

Techniques Used: Sonication

Effect of heat and ciprofloxacin on the biofilm. Biofilms were incubated at 37 °C with the indicated ciprofloxacin concentration for 24 h before enumeration. 4 h into the incubation they were heat shocked for the indicated time and temperature (in some cases at 37 °C as controls). Error bars indicate SD (n = 12). Asterisks indicate trials that were statistically different from the corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p
Figure Legend Snippet: Effect of heat and ciprofloxacin on the biofilm. Biofilms were incubated at 37 °C with the indicated ciprofloxacin concentration for 24 h before enumeration. 4 h into the incubation they were heat shocked for the indicated time and temperature (in some cases at 37 °C as controls). Error bars indicate SD (n = 12). Asterisks indicate trials that were statistically different from the corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p

Techniques Used: Incubation, Concentration Assay

Effect of heat and erythromycin on the biofilm. Biofilms were incubated at 37 °C with the indicated erythromycin concentrations for 24 h before enumeration. 4 h into this incubation the biofilms were heat shocked for the indicated time and temperature (in some cases at 37 °C as controls). Error bars indicate the SD (n = 12). Asterisks indicate trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p
Figure Legend Snippet: Effect of heat and erythromycin on the biofilm. Biofilms were incubated at 37 °C with the indicated erythromycin concentrations for 24 h before enumeration. 4 h into this incubation the biofilms were heat shocked for the indicated time and temperature (in some cases at 37 °C as controls). Error bars indicate the SD (n = 12). Asterisks indicate trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p

Techniques Used: Incubation, Concentration Assay

Effect of tobramycin on the planktonic bacteria dispersed from the biofilm. Antibiotic efficacy against the planktonic bacteria dispersed from the heat shocked biofilms was measured by optical density. Error bars indicate the SD (n = 12). A baseline correction of 0.8 based on negative controls was applied. Biofilms which died before enumeration also showed no surviving planktonic bacteria. However, planktonic bacteria from surviving biofilms were also eliminated at tobramycin concentrations of 2 μg mland above. Asterisks indicated trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p
Figure Legend Snippet: Effect of tobramycin on the planktonic bacteria dispersed from the biofilm. Antibiotic efficacy against the planktonic bacteria dispersed from the heat shocked biofilms was measured by optical density. Error bars indicate the SD (n = 12). A baseline correction of 0.8 based on negative controls was applied. Biofilms which died before enumeration also showed no surviving planktonic bacteria. However, planktonic bacteria from surviving biofilms were also eliminated at tobramycin concentrations of 2 μg mland above. Asterisks indicated trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p

Techniques Used: Concentration Assay

Antibiotic effect on biofilms. The ciprofloxacin and tobramycin followed a power-law decrease for a discrete concentration range before reaching an asymptote, with ciprofloxacin having a larger effect on the biofilms than tobramycin. Erythromycin showed no statistical differences for each concentration tested.
Figure Legend Snippet: Antibiotic effect on biofilms. The ciprofloxacin and tobramycin followed a power-law decrease for a discrete concentration range before reaching an asymptote, with ciprofloxacin having a larger effect on the biofilms than tobramycin. Erythromycin showed no statistical differences for each concentration tested.

Techniques Used: Concentration Assay

Effect of erythromycin on the planktonic bacteria dispersed from the biofilm. Antibiotic efficacy against the planktonic bacteria dispersed from the heat shocked biofilms was measured by optical density. Error bars indicate the SD (n = 12). A baseline correction of 0.8 based on negative controls was applied. Biofilms which died out before enumeration also showed no surviving planktonic bacteria. However, planktonic bacteria from surviving biofilms were also reduced at erythromycin concentrations of 64 μg ml and above. Asterisks indicated trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p
Figure Legend Snippet: Effect of erythromycin on the planktonic bacteria dispersed from the biofilm. Antibiotic efficacy against the planktonic bacteria dispersed from the heat shocked biofilms was measured by optical density. Error bars indicate the SD (n = 12). A baseline correction of 0.8 based on negative controls was applied. Biofilms which died out before enumeration also showed no surviving planktonic bacteria. However, planktonic bacteria from surviving biofilms were also reduced at erythromycin concentrations of 64 μg ml and above. Asterisks indicated trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p

Techniques Used: Concentration Assay

Effect of ciprofloxacin on the planktonic bacteria dispersed from the biofilm. Antibiotic efficacy against the planktonic bacteria dispersed from the heat shocked biofilms was measured by optical density. Error bars indicate the SD (n = 12). A baseline correction of 0.8 based on negative controls was applied. Biofilms which died before enumeration also showed no surviving planktonic bacteria. However, planktonic bacteria from surviving biofilms were also eliminated at ciprofloxacin concentrations of 1 μg ml −1 and above. Asterisks indicate trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p
Figure Legend Snippet: Effect of ciprofloxacin on the planktonic bacteria dispersed from the biofilm. Antibiotic efficacy against the planktonic bacteria dispersed from the heat shocked biofilms was measured by optical density. Error bars indicate the SD (n = 12). A baseline correction of 0.8 based on negative controls was applied. Biofilms which died before enumeration also showed no surviving planktonic bacteria. However, planktonic bacteria from surviving biofilms were also eliminated at ciprofloxacin concentrations of 1 μg ml −1 and above. Asterisks indicate trials that were statistically different from their corresponding controls (the 37 °C within that concentration group) as determined by two-way ANOVA (p

Techniques Used: Concentration Assay

2) Product Images from "Flow cytometry combined with viSNE for the analysis of microbial biofilms and detection of microplastics"

Article Title: Flow cytometry combined with viSNE for the analysis of microbial biofilms and detection of microplastics

Journal: Nature Communications

doi: 10.1038/ncomms11587

Tracking and quantification of biofilm community structure changes after temperature increase. ( a ) viSNE submaps ( Fig. 2a ) belonging to days 0, 7, 14 and 21 post temperature increase. More colour-intense regions of the submaps depict regions of higher cell density. Similarity analysis of technical replicates (that is, three per sample), biological replicates (that is, five independent microcosms) and time points indicated that the detected changes in community structure were governed by time point and not biological or technical noise ( Supplementary Fig. 10 ). ( b ) Quantification of subpopulations defined in Fig. 2b , pooled from five biological replicates for each time point after temperature increase (all biological replicates are depicted in Supplementary Fig. 11 ). Statistical analysis of subpopulation sizes is available in Supplementary Fig. 9 .
Figure Legend Snippet: Tracking and quantification of biofilm community structure changes after temperature increase. ( a ) viSNE submaps ( Fig. 2a ) belonging to days 0, 7, 14 and 21 post temperature increase. More colour-intense regions of the submaps depict regions of higher cell density. Similarity analysis of technical replicates (that is, three per sample), biological replicates (that is, five independent microcosms) and time points indicated that the detected changes in community structure were governed by time point and not biological or technical noise ( Supplementary Fig. 10 ). ( b ) Quantification of subpopulations defined in Fig. 2b , pooled from five biological replicates for each time point after temperature increase (all biological replicates are depicted in Supplementary Fig. 11 ). Statistical analysis of subpopulation sizes is available in Supplementary Fig. 9 .

Techniques Used:

Categorizing subpopulations in temperature-stressed stream biofilms. ( a ) Stream biofilms were assessed by FC directly after transfer to higher temperature, after 1, 2 and 3 weeks and the acquired data was altogether mapped by viSNE. viSNE maps are shown in single colour, with each point in the viSNE map representing a single cell from the biofilms, or coloured according to FS and fluorescence intensity at specific wavelengths (nm) measured by FC (full set of wavelengths displayed in Supplementary Fig. 6 ). ( b ) Subpopulations (LA1–LA15) categorized based on the viSNE map, optical scatter and fluorescence intensities ( a ) ( Supplementary Fig. 6 ). A fraction of the particles (4.5–5.8%) was not categorized due to lack of distinct properties. Comparison of subpopulation properties with data acquired from reference species and pigment-bleached reference samples allowed for assigning subpopulations to types of organisms and potentially decaying cells ( Supplementary Fig. 8 ).
Figure Legend Snippet: Categorizing subpopulations in temperature-stressed stream biofilms. ( a ) Stream biofilms were assessed by FC directly after transfer to higher temperature, after 1, 2 and 3 weeks and the acquired data was altogether mapped by viSNE. viSNE maps are shown in single colour, with each point in the viSNE map representing a single cell from the biofilms, or coloured according to FS and fluorescence intensity at specific wavelengths (nm) measured by FC (full set of wavelengths displayed in Supplementary Fig. 6 ). ( b ) Subpopulations (LA1–LA15) categorized based on the viSNE map, optical scatter and fluorescence intensities ( a ) ( Supplementary Fig. 6 ). A fraction of the particles (4.5–5.8%) was not categorized due to lack of distinct properties. Comparison of subpopulation properties with data acquired from reference species and pigment-bleached reference samples allowed for assigning subpopulations to types of organisms and potentially decaying cells ( Supplementary Fig. 8 ).

Techniques Used: Fluorescence

Tracking biofilm community structure changes along a stream. ( a ) viSNE submaps belonging to six sites (A–F) along the stream Mönchaltorfer Aa. More colour-intense regions of the submaps depict regions of higher cell or particle density. Similarity analysis of technical replicates (that is, three per sample), biological replicates (that is, 3 stones per site) and sites indicated that the detected changes in community structure were governed by site and not biological or technical noise ( Supplementary Fig. 17 ). The sites are characterized in Supplementary Tables 4–6 . Site A is at the spring of the stream in the forest, site B is in an unshaded stretch, site C is shaded but in the straightened section of the stream-like sites D–F, which are additionally influenced by waste-water treatment plant effluents, site D being situated immediately downstream a treatment plant. ( b ) Subpopulations are defined in Fig. 4b , pooled from three biological replicates taken from each site (all biological replicates are depicted in Supplementary Fig. 18 ). Statistical analysis of subpopulation sizes is available in Supplementary Fig. 16 . ( c ) Biplots of the redundancy analysis (RDA) based on the fraction of cells/particles in the subpopulations in Fig. 4b constrained by forward selected field physico-chemical parameters ( Supplementary Tables 4,6 ). Dots/grey tones: specific sampling sites. Dispersion of standard error of the weighted scores of sampling sites are shown as ellipses in the respective grey tone (confidence limit=0.95). Centroids of the subpopulations (MA1a–MA10) are given. Significantly tested model variables are depicted (*** P
Figure Legend Snippet: Tracking biofilm community structure changes along a stream. ( a ) viSNE submaps belonging to six sites (A–F) along the stream Mönchaltorfer Aa. More colour-intense regions of the submaps depict regions of higher cell or particle density. Similarity analysis of technical replicates (that is, three per sample), biological replicates (that is, 3 stones per site) and sites indicated that the detected changes in community structure were governed by site and not biological or technical noise ( Supplementary Fig. 17 ). The sites are characterized in Supplementary Tables 4–6 . Site A is at the spring of the stream in the forest, site B is in an unshaded stretch, site C is shaded but in the straightened section of the stream-like sites D–F, which are additionally influenced by waste-water treatment plant effluents, site D being situated immediately downstream a treatment plant. ( b ) Subpopulations are defined in Fig. 4b , pooled from three biological replicates taken from each site (all biological replicates are depicted in Supplementary Fig. 18 ). Statistical analysis of subpopulation sizes is available in Supplementary Fig. 16 . ( c ) Biplots of the redundancy analysis (RDA) based on the fraction of cells/particles in the subpopulations in Fig. 4b constrained by forward selected field physico-chemical parameters ( Supplementary Tables 4,6 ). Dots/grey tones: specific sampling sites. Dispersion of standard error of the weighted scores of sampling sites are shown as ellipses in the respective grey tone (confidence limit=0.95). Centroids of the subpopulations (MA1a–MA10) are given. Significantly tested model variables are depicted (*** P

Techniques Used: Sampling

Categorizing subpopulations in stream biofilms sampled in the field. ( a ) Stream biofilms were assessed by FC after sampling at six sites along the stream Mönchaltorfer Aa and altogether mapped by viSNE. viSNE maps are shown in single colour, with each point in the viSNE map representing a single cell or particle from the biofilms or coloured according to FS and fluorescence intensity at specific wavelengths (nm) measured by FC (full set displayed in Supplementary Fig. 13 ). ( b ) Subpopulations (MA 1–10) categorized based on the viSNE map and optical scatter and fluorescence intensities ( a ) ( Supplementary Fig. 13 ). Some cells (range 2.7–11.2 %) were not categorized due to lack of distinct properties. Comparison of subpopulation properties with data acquired from reference species and pigment-bleached reference samples ( Supplementary Fig. 15 ) allowed for assigning subpopulations to types of organisms and potentially decaying cells.
Figure Legend Snippet: Categorizing subpopulations in stream biofilms sampled in the field. ( a ) Stream biofilms were assessed by FC after sampling at six sites along the stream Mönchaltorfer Aa and altogether mapped by viSNE. viSNE maps are shown in single colour, with each point in the viSNE map representing a single cell or particle from the biofilms or coloured according to FS and fluorescence intensity at specific wavelengths (nm) measured by FC (full set displayed in Supplementary Fig. 13 ). ( b ) Subpopulations (MA 1–10) categorized based on the viSNE map and optical scatter and fluorescence intensities ( a ) ( Supplementary Fig. 13 ). Some cells (range 2.7–11.2 %) were not categorized due to lack of distinct properties. Comparison of subpopulation properties with data acquired from reference species and pigment-bleached reference samples ( Supplementary Fig. 15 ) allowed for assigning subpopulations to types of organisms and potentially decaying cells.

Techniques Used: Sampling, Fluorescence

Detection of microplastics in stream biofilms. ( a ) viSNE map shown in Fig. 4a with circled hypothetical microplastic cluster. ( b ) SEM image of a potential microplastic particle isolated from site D. Scale bar, 10 μm. ( c ) SEM image of polystyrene beads isolated from a spiked sample. Scale bar, 50 μm. The squares in the top right corner of the SEM images ( b , c ) with separate scale bars, 1 μm, depict the region scanned for EDS analysis. ( d ) EDS spectrum of a potential microplastic particle isolated from site D. ( e ) EDS spectrum of polystyrene beads isolated from a spiked sample.
Figure Legend Snippet: Detection of microplastics in stream biofilms. ( a ) viSNE map shown in Fig. 4a with circled hypothetical microplastic cluster. ( b ) SEM image of a potential microplastic particle isolated from site D. Scale bar, 10 μm. ( c ) SEM image of polystyrene beads isolated from a spiked sample. Scale bar, 50 μm. The squares in the top right corner of the SEM images ( b , c ) with separate scale bars, 1 μm, depict the region scanned for EDS analysis. ( d ) EDS spectrum of a potential microplastic particle isolated from site D. ( e ) EDS spectrum of polystyrene beads isolated from a spiked sample.

Techniques Used: Isolation

3) Product Images from "Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium"

Article Title: Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00602-19

Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P
Figure Legend Snippet: Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P

Techniques Used: Activity Assay, Fluorescence

Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P
Figure Legend Snippet: Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P

Techniques Used: Activity Assay, Fluorescence, Incubation

Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.
Figure Legend Snippet: Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.

Techniques Used:

4) Product Images from "A Combined Pharmacodynamic Quantitative and Qualitative Model Reveals the Potent Activity of Daptomycin and Delafloxacin against Staphylococcus aureus Biofilms"

Article Title: A Combined Pharmacodynamic Quantitative and Qualitative Model Reveals the Potent Activity of Daptomycin and Delafloxacin against Staphylococcus aureus Biofilms

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00181-13

Characterization of the biofilm model with MSSA ATCC 25923. (Left) Evolution over time of the crystal violet absorbance (as a marker of biofilm production) and of resorufin fluorescence (as a marker of bacterial viability) for an initial inoculum with
Figure Legend Snippet: Characterization of the biofilm model with MSSA ATCC 25923. (Left) Evolution over time of the crystal violet absorbance (as a marker of biofilm production) and of resorufin fluorescence (as a marker of bacterial viability) for an initial inoculum with

Techniques Used: Marker, Fluorescence

Antibiotic activities against 6-h S. aureus biofilms.
Figure Legend Snippet: Antibiotic activities against 6-h S. aureus biofilms.

Techniques Used:

(Left and middle) Three-dimensional images from confocal laser scanning microscopy of 24-h biofilms of MSSA ATCC 25923 (left) and MRSA ATCC 33591 (middle) under control conditions or after exposure to selected antibiotics at 32 times their MICs for 48
Figure Legend Snippet: (Left and middle) Three-dimensional images from confocal laser scanning microscopy of 24-h biofilms of MSSA ATCC 25923 (left) and MRSA ATCC 33591 (middle) under control conditions or after exposure to selected antibiotics at 32 times their MICs for 48

Techniques Used: Confocal Laser Scanning Microscopy

Concentration-response activities of antibiotics against 6-h biofilms of MSSA ATCC 25923 (left) or MRSA ATCC 33591 (right). The 6-h biofilms were incubated with increasing concentrations of antibiotics (shown on the x axis) for 24 h. The ordinate shows
Figure Legend Snippet: Concentration-response activities of antibiotics against 6-h biofilms of MSSA ATCC 25923 (left) or MRSA ATCC 33591 (right). The 6-h biofilms were incubated with increasing concentrations of antibiotics (shown on the x axis) for 24 h. The ordinate shows

Techniques Used: Concentration Assay, Incubation

Concentration-response activities of antibiotics against 24-h biofilms of MSSA ATCC 25923 (left) or MRSA ATCC 33591 (right). The 24-h biofilms were incubated with increasing concentrations of antibiotics (shown on the x axis) for 48 h. The ordinate shows
Figure Legend Snippet: Concentration-response activities of antibiotics against 24-h biofilms of MSSA ATCC 25923 (left) or MRSA ATCC 33591 (right). The 24-h biofilms were incubated with increasing concentrations of antibiotics (shown on the x axis) for 48 h. The ordinate shows

Techniques Used: Concentration Assay, Incubation

(Left) Three-dimensional images from confocal laser scanning microscopy of 24-h biofilms of MSSA ATCC 25923 (top) or MRSA ATCC 33591 (bottom) under control conditions or after exposure to delafloxacin or daptomycin at 8 and 16 times the respective MIC
Figure Legend Snippet: (Left) Three-dimensional images from confocal laser scanning microscopy of 24-h biofilms of MSSA ATCC 25923 (top) or MRSA ATCC 33591 (bottom) under control conditions or after exposure to delafloxacin or daptomycin at 8 and 16 times the respective MIC

Techniques Used: Confocal Laser Scanning Microscopy

Antibiotic activity against 24-h S. aureus biofilms.
Figure Legend Snippet: Antibiotic activity against 24-h S. aureus biofilms.

Techniques Used: Activity Assay

5) Product Images from "Silver Oxynitrate, an Unexplored Silver Compound with Antimicrobial and Antibiofilm Activity"

Article Title: Silver Oxynitrate, an Unexplored Silver Compound with Antimicrobial and Antibiofilm Activity

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.05177-14

Ag 7 NO 11 eradicates established biofilms at lower concentrations than those of other metal compounds. The MBEC device was inoculated with E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923). Biofilms were established following a 24-h incubation.
Figure Legend Snippet: Ag 7 NO 11 eradicates established biofilms at lower concentrations than those of other metal compounds. The MBEC device was inoculated with E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923). Biofilms were established following a 24-h incubation.

Techniques Used: Incubation

Ag 7 NO 11 is more efficacious for eradicating mature biofilms of E. coli and S. aureus . E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923) biofilms were cultivated by using the MBEC device. Established biofilms were formed following incubation
Figure Legend Snippet: Ag 7 NO 11 is more efficacious for eradicating mature biofilms of E. coli and S. aureus . E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923) biofilms were cultivated by using the MBEC device. Established biofilms were formed following incubation

Techniques Used: Incubation

Ag 7 NO 11 has antimicrobial and antibiofilm activity against a MRSA strain (USA300). MRSA biofilms were cultivated by using the MBEC device. (A and B) The MBC for the planktonic population (A) and the MBEC for the biofilm population (B) were determined
Figure Legend Snippet: Ag 7 NO 11 has antimicrobial and antibiofilm activity against a MRSA strain (USA300). MRSA biofilms were cultivated by using the MBEC device. (A and B) The MBC for the planktonic population (A) and the MBEC for the biofilm population (B) were determined

Techniques Used: Activity Assay

Ag 7 NO 11 has antimicrobial and antibiofilm activity against UPEC (CFT703). UPEC biofilms were cultivated by using the MBEC device. (A and B) The MBC for the planktonic population (A) and the MBEC for the biofilm population (B) were determined after a 4-h
Figure Legend Snippet: Ag 7 NO 11 has antimicrobial and antibiofilm activity against UPEC (CFT703). UPEC biofilms were cultivated by using the MBEC device. (A and B) The MBC for the planktonic population (A) and the MBEC for the biofilm population (B) were determined after a 4-h

Techniques Used: Activity Assay

Ag 7 NO 11 reduces biofilm quantity. Biofilms of E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923) were established in the MBEC device for 24 h. The established biofilms were then exposed to 0 μM (A), 5 μM (B), and 12.5 μM
Figure Legend Snippet: Ag 7 NO 11 reduces biofilm quantity. Biofilms of E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923) were established in the MBEC device for 24 h. The established biofilms were then exposed to 0 μM (A), 5 μM (B), and 12.5 μM

Techniques Used:

Ag 7 NO 11 prevents biofilm formation at lower concentrations than those of other metal compounds. The MBEC device was inoculated with E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923) concurrently with serial dilutions (2-fold) of Ag 2 O,
Figure Legend Snippet: Ag 7 NO 11 prevents biofilm formation at lower concentrations than those of other metal compounds. The MBEC device was inoculated with E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923) concurrently with serial dilutions (2-fold) of Ag 2 O,

Techniques Used:

Ag 7 NO 11 has antimicrobial and antibiofilm activity against an FQRP isolate. FQRP biofilms were cultivated by using the MBEC device. (A and B) The MBC for the planktonic population (A) and the MBEC for the biofilm population (B) were determined after a
Figure Legend Snippet: Ag 7 NO 11 has antimicrobial and antibiofilm activity against an FQRP isolate. FQRP biofilms were cultivated by using the MBEC device. (A and B) The MBC for the planktonic population (A) and the MBEC for the biofilm population (B) were determined after a

Techniques Used: Activity Assay

Ag 7 NO 11 reduces biofilm biomass. Biofilms of E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923) were established in the MBEC device for 24 h. The biofilms were then exposed to various concentrations of CuSO 4 (A), AgNO 3 (B), and Ag 7 NO 11
Figure Legend Snippet: Ag 7 NO 11 reduces biofilm biomass. Biofilms of E. coli (JM109), P. aeruginosa (PAO1), and S. aureus (ATCC 25923) were established in the MBEC device for 24 h. The biofilms were then exposed to various concentrations of CuSO 4 (A), AgNO 3 (B), and Ag 7 NO 11

Techniques Used:

6) Product Images from "Enhanced Surface Colonization by Escherichia coli O157:H7 in Biofilms Formed by an Acinetobacter calcoaceticus Isolate from Meat-Processing Environments ▿ Isolate from Meat-Processing Environments ▿ †"

Article Title: Enhanced Surface Colonization by Escherichia coli O157:H7 in Biofilms Formed by an Acinetobacter calcoaceticus Isolate from Meat-Processing Environments ▿ Isolate from Meat-Processing Environments ▿ †

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.02707-09

Biovolume of A. calcoaceticus and E. coli O157:H7 biofilm development after 24, 48, and 72 h of growth under dynamic conditions. A. calcoaceticus in monospecies biofilms is represented by the symbol □, A. calcoaceticus in dual-species biofilms
Figure Legend Snippet: Biovolume of A. calcoaceticus and E. coli O157:H7 biofilm development after 24, 48, and 72 h of growth under dynamic conditions. A. calcoaceticus in monospecies biofilms is represented by the symbol □, A. calcoaceticus in dual-species biofilms

Techniques Used:

Structural development of A. calcoaceticus and E. coli in mono- and dual-species biofilms under dynamic conditions. (A) Representative biofilms of A. calcoaceticus and pGFP-uv-tagged E. coli O157:H7 grown in flow cells using Luria-Bertani broth as a growth
Figure Legend Snippet: Structural development of A. calcoaceticus and E. coli in mono- and dual-species biofilms under dynamic conditions. (A) Representative biofilms of A. calcoaceticus and pGFP-uv-tagged E. coli O157:H7 grown in flow cells using Luria-Bertani broth as a growth

Techniques Used: Flow Cytometry

7) Product Images from "Pseudomonas aeruginosa Leucine Aminopeptidase Influences Early Biofilm Composition and Structure via Vesicle-Associated Antibiofilm Activity"

Article Title: Pseudomonas aeruginosa Leucine Aminopeptidase Influences Early Biofilm Composition and Structure via Vesicle-Associated Antibiofilm Activity

Journal: mBio

doi: 10.1128/mBio.02548-19

PaAP deletion modifies pellicle cellular biomass and matrix composition, and WT OMV treatment complements these phenotypes. Untreated: (A) S470 WT or S470 ΔPaAP was grown in 35-mm dishes for 6 h, and pellicle biofilms were quantified by crystal violet staining (OD 495 ). (B and C) Pellicles formed by S470 WT or S470 ΔPaAP after 6 h of growth were live/dead stained (B) and imaged by confocal microscopy (C), and their biomass was calculated. (D and E) Pellicles were stained for matrix Psl (D) and total protein (E), and the results were quantified and normalized to cellular volume and Psl volume, respectively. Treated: (A and C to E) ΔPaAP pellicles were treated at 1 hpi with S470 WT or ΔPaAP OMVs and imaged at 5 hpi. Untreated WT and treated ΔPaAP pellicles were stained as described above and quantified, and values were compared to untreated ΔPaAP pellicles. *, P
Figure Legend Snippet: PaAP deletion modifies pellicle cellular biomass and matrix composition, and WT OMV treatment complements these phenotypes. Untreated: (A) S470 WT or S470 ΔPaAP was grown in 35-mm dishes for 6 h, and pellicle biofilms were quantified by crystal violet staining (OD 495 ). (B and C) Pellicles formed by S470 WT or S470 ΔPaAP after 6 h of growth were live/dead stained (B) and imaged by confocal microscopy (C), and their biomass was calculated. (D and E) Pellicles were stained for matrix Psl (D) and total protein (E), and the results were quantified and normalized to cellular volume and Psl volume, respectively. Treated: (A and C to E) ΔPaAP pellicles were treated at 1 hpi with S470 WT or ΔPaAP OMVs and imaged at 5 hpi. Untreated WT and treated ΔPaAP pellicles were stained as described above and quantified, and values were compared to untreated ΔPaAP pellicles. *, P

Techniques Used: Staining, Confocal Microscopy

PaAP expression promotes Psl production in coculture biofilms. (A) Bacterial biofilms cocultured on A549 cells were imaged by confocal microscopy at indicated time points. (S470 WT, dark gray; S470 ΔPaAP, light gray). (B) Total protein was extracted from S470 WT and ΔPaAP cocultures at indicated hours postinoculation and precipitated, and PaAP was detected in the samples using Western blotting. Mwt, 50-kDa standard; PaAP, purified PaAP control. (C and D) Bacterial biofilms (cyan) were cocultured on A549 cells for 5 h, stained with HHA-FITC lectin (magenta), and imaged by confocal microscopy (C), and the ratio of Psl/cellular biomass was calculated (D). (E) At 3 hpi, S470 WT (solid line) and ΔPaAP (dashed line) cocultures were treated with the indicated concentrations of colistin. Survival indicates the percentage of biomass remaining attached to the host cell substrate. For all experiments, representative results are shown. *, P
Figure Legend Snippet: PaAP expression promotes Psl production in coculture biofilms. (A) Bacterial biofilms cocultured on A549 cells were imaged by confocal microscopy at indicated time points. (S470 WT, dark gray; S470 ΔPaAP, light gray). (B) Total protein was extracted from S470 WT and ΔPaAP cocultures at indicated hours postinoculation and precipitated, and PaAP was detected in the samples using Western blotting. Mwt, 50-kDa standard; PaAP, purified PaAP control. (C and D) Bacterial biofilms (cyan) were cocultured on A549 cells for 5 h, stained with HHA-FITC lectin (magenta), and imaged by confocal microscopy (C), and the ratio of Psl/cellular biomass was calculated (D). (E) At 3 hpi, S470 WT (solid line) and ΔPaAP (dashed line) cocultures were treated with the indicated concentrations of colistin. Survival indicates the percentage of biomass remaining attached to the host cell substrate. For all experiments, representative results are shown. *, P

Techniques Used: Expressing, Confocal Microscopy, Western Blot, Purification, Staining

Summary of the effect of PaAP + OMVs on early biofilm development. During WT growth, bacterial microcolonies secrete PaAP + OMVs with increased endogenous protease activity. This leads to increased cell detachment from the colony structure and allows for increased matrix Psl polysaccharide production and protection for the cells against antibiotic treatment. Orange squares, A549 epithelial cells; green cells, P. aeruginosa cells; red circles, PaAP + OMVs; blue circles, ΔPaAP OMVs; green haze, matrix Psl. Darker matrix color indicates a higher concentration of protein.
Figure Legend Snippet: Summary of the effect of PaAP + OMVs on early biofilm development. During WT growth, bacterial microcolonies secrete PaAP + OMVs with increased endogenous protease activity. This leads to increased cell detachment from the colony structure and allows for increased matrix Psl polysaccharide production and protection for the cells against antibiotic treatment. Orange squares, A549 epithelial cells; green cells, P. aeruginosa cells; red circles, PaAP + OMVs; blue circles, ΔPaAP OMVs; green haze, matrix Psl. Darker matrix color indicates a higher concentration of protein.

Techniques Used: Activity Assay, Concentration Assay

PaAP + OMVs cause biofilm detachment in both pseudomonads and nonpseudomonads. (A and B) OMVs from WT or ΔPaAP cultures of the indicated P. aeruginosa strains were added to S470 ΔPaAP biofilm cocultures at 4.5 hpi, 30 min prior to imaging (A), and these results were quantified (B). The biomasses of treated biofilms and the S470 WT untreated controls were quantified and were compared with untreated S470 ΔPaAP biofilms. (C and D) K. pneumoniae cocultures were treated with S470 WT or ΔPaAP vesicles at 4.5 hpi, and biofilm formation was assessed by Congo red staining at 5 hpi (C) and was quantified (D). The biomasses of treated K. pneumoniae biofilms were quantified and were compared with untreated biofilms. For all experiments, representative results are shown. *, P
Figure Legend Snippet: PaAP + OMVs cause biofilm detachment in both pseudomonads and nonpseudomonads. (A and B) OMVs from WT or ΔPaAP cultures of the indicated P. aeruginosa strains were added to S470 ΔPaAP biofilm cocultures at 4.5 hpi, 30 min prior to imaging (A), and these results were quantified (B). The biomasses of treated biofilms and the S470 WT untreated controls were quantified and were compared with untreated S470 ΔPaAP biofilms. (C and D) K. pneumoniae cocultures were treated with S470 WT or ΔPaAP vesicles at 4.5 hpi, and biofilm formation was assessed by Congo red staining at 5 hpi (C) and was quantified (D). The biomasses of treated K. pneumoniae biofilms were quantified and were compared with untreated biofilms. For all experiments, representative results are shown. *, P

Techniques Used: Imaging, Staining

PaAP increases protease activity in OMVs, leading to cell detachment from biofilm microcolonies. (A) S470 ΔPaAP cocultures (solid line) were challenged with colistin at 3 hpi and examined for remaining biomass at 5 hpi. Additional S470 ΔPaAP cocultures were treated with S470 WT OMVs at 4.5 hpi, and the detached cells (dotted line) were collected at 5 hpi and challenged with colistin for 2 h. These samples were compared to planktonically grown S470 ΔPaAP cells (dashed line) challenged with colistin at 3 hpi with live/dead assessment at 5 hpi. (B) S470 ΔPaAP biofilm cocultures were treated with OMVs isolated from PA14 WT and the indicated PA14 transposon insertion mutants at 4.5 hpi, and the biomass was quantified at 5 hpi. The biomasses of treated biofilms and the S470 WT untreated control were quantified, and treated samples were compared to biofilms treated with S470 WT OMVs. (C) Protease activity was measured in the indicated S470 WT and ΔPaAP OMV preparations with and without protease inhibitor cocktail (PI) and for rPaAP. (D) S470 ΔPaAP biofilm cocultures were treated with the indicated S470 WT and ΔPaAP OMVs with and without PI at 4.5 hpi, and the resulting biomass was quantified at 5 hpi. The biomasses of treated biofilms, the S470 WT untreated control, and the S470 ΔPaAP biofilm incubated with PI were quantified, and samples were compared to the corresponding samples without PI. For microscopy-based experiments, representative results are shown. For all other experiments, n = 3. *, P
Figure Legend Snippet: PaAP increases protease activity in OMVs, leading to cell detachment from biofilm microcolonies. (A) S470 ΔPaAP cocultures (solid line) were challenged with colistin at 3 hpi and examined for remaining biomass at 5 hpi. Additional S470 ΔPaAP cocultures were treated with S470 WT OMVs at 4.5 hpi, and the detached cells (dotted line) were collected at 5 hpi and challenged with colistin for 2 h. These samples were compared to planktonically grown S470 ΔPaAP cells (dashed line) challenged with colistin at 3 hpi with live/dead assessment at 5 hpi. (B) S470 ΔPaAP biofilm cocultures were treated with OMVs isolated from PA14 WT and the indicated PA14 transposon insertion mutants at 4.5 hpi, and the biomass was quantified at 5 hpi. The biomasses of treated biofilms and the S470 WT untreated control were quantified, and treated samples were compared to biofilms treated with S470 WT OMVs. (C) Protease activity was measured in the indicated S470 WT and ΔPaAP OMV preparations with and without protease inhibitor cocktail (PI) and for rPaAP. (D) S470 ΔPaAP biofilm cocultures were treated with the indicated S470 WT and ΔPaAP OMVs with and without PI at 4.5 hpi, and the resulting biomass was quantified at 5 hpi. The biomasses of treated biofilms, the S470 WT untreated control, and the S470 ΔPaAP biofilm incubated with PI were quantified, and samples were compared to the corresponding samples without PI. For microscopy-based experiments, representative results are shown. For all other experiments, n = 3. *, P

Techniques Used: Activity Assay, Isolation, Protease Inhibitor, Incubation, Microscopy

Deletion of the PaAP aminopeptidase increased the density, biomass, and organization of bacterial biofilms on host cells. (A) S470 and PAO1 WT and ΔPaAP strains were inoculated onto confluent A549 cell layers, and images were taken at ×10 magnification at 5 hpi. (B and C) The images from panel A were quantified. (D and E) For cocultures as described for panel A, images of microcolonies were taken using ×100 magnification at 5 hpi (D) and quantified (E). For all experiments, representative results are shown. Quantified bacterial biomasses in each set of cocultures were compared. *, P
Figure Legend Snippet: Deletion of the PaAP aminopeptidase increased the density, biomass, and organization of bacterial biofilms on host cells. (A) S470 and PAO1 WT and ΔPaAP strains were inoculated onto confluent A549 cell layers, and images were taken at ×10 magnification at 5 hpi. (B and C) The images from panel A were quantified. (D and E) For cocultures as described for panel A, images of microcolonies were taken using ×100 magnification at 5 hpi (D) and quantified (E). For all experiments, representative results are shown. Quantified bacterial biomasses in each set of cocultures were compared. *, P

Techniques Used:

Addition of PaAP + OMVs inhibits formation of P. aeruginosa coculture biofilms. (A) Cell-free supernatants (CFS) from S470 WT and S470 ΔPaAP cultures were TCA precipitated, and PaAP was detected by immunoblotting after separation of samples by SDS-PAGE. The relative migration of rPaAP and the 50-kDa molecular weight standard (Mwt) are shown. Samples were run on the same gel, and an intervening lane was excised in this figure. (B to D) S470 ΔPaAP biofilms cocultured with A549 cells were treated with CFS S470 WT or ΔPaAP at 1 hpi (B), with either light (OMVs) or dense (heavy) fractions from S470 WT or S470 ΔPaAP CFS (C), or with rPaAP, rPaAP with ΔPaAP OMVs, or sPaAP with ΔPaAP OMVs (D). The biomasses of these treated biofilms and the S470 WT untreated control were quantified at 5 hpi and were compared with untreated S470 ΔPaAP biofilms. *, P
Figure Legend Snippet: Addition of PaAP + OMVs inhibits formation of P. aeruginosa coculture biofilms. (A) Cell-free supernatants (CFS) from S470 WT and S470 ΔPaAP cultures were TCA precipitated, and PaAP was detected by immunoblotting after separation of samples by SDS-PAGE. The relative migration of rPaAP and the 50-kDa molecular weight standard (Mwt) are shown. Samples were run on the same gel, and an intervening lane was excised in this figure. (B to D) S470 ΔPaAP biofilms cocultured with A549 cells were treated with CFS S470 WT or ΔPaAP at 1 hpi (B), with either light (OMVs) or dense (heavy) fractions from S470 WT or S470 ΔPaAP CFS (C), or with rPaAP, rPaAP with ΔPaAP OMVs, or sPaAP with ΔPaAP OMVs (D). The biomasses of these treated biofilms and the S470 WT untreated control were quantified at 5 hpi and were compared with untreated S470 ΔPaAP biofilms. *, P

Techniques Used: SDS Page, Migration, Molecular Weight

8) Product Images from "Spermine inhibits Vibrio cholerae biofilm formation through the NspS–MbaA polyamine signaling system"

Article Title: Spermine inhibits Vibrio cholerae biofilm formation through the NspS–MbaA polyamine signaling system

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M117.801068

Spermine inhibits vpsL gene transcription in a NspS—MbaA-dependent manner. Static cultures were grown for 20 h with the indicated concentration of spermine ( Spm ) at 27 °C. Planktonic cells and biofilms were homogenized by disruption with glass beads and vortexing. Cells were pelleted and lysed in Z-buffer. Lysates were incubated with the colorimetric β-galactosidase substrate ortho -nitrophenyl-β-galactoside, and Miller units were calculated using A 415 of reaction mixtures. Shown are the averages and S.D. ( error bars ) of three experiments with three technical replicates each. Means not followed by the same letter are statistically different at p ≤ 0.05 from the untreated samples within a given strain, as determined by one-way ANOVA using SigmaPlot version 12.5 ( NS , not significant).
Figure Legend Snippet: Spermine inhibits vpsL gene transcription in a NspS—MbaA-dependent manner. Static cultures were grown for 20 h with the indicated concentration of spermine ( Spm ) at 27 °C. Planktonic cells and biofilms were homogenized by disruption with glass beads and vortexing. Cells were pelleted and lysed in Z-buffer. Lysates were incubated with the colorimetric β-galactosidase substrate ortho -nitrophenyl-β-galactoside, and Miller units were calculated using A 415 of reaction mixtures. Shown are the averages and S.D. ( error bars ) of three experiments with three technical replicates each. Means not followed by the same letter are statistically different at p ≤ 0.05 from the untreated samples within a given strain, as determined by one-way ANOVA using SigmaPlot version 12.5 ( NS , not significant).

Techniques Used: Concentration Assay, Incubation

Spermine inhibits V. cholerae biofilms in an NspS/MbaA-dependent manner. Static cultures were incubated for 20 h at 27 °C in the LB only or with varying spermine concentrations. Biofilms were washed, disrupted, and measured at A 595 ( Spm , spermine). Shown are averages and S.D. ( error bars ) of three separate experiments with three technical replicates each. Means not followed by the same letter are statistically different at p ≤ 0.05 from the untreated samples within a given strain, as determined by one-way ANOVA using SigmaPlot version 12.5 ( NS , not significant).
Figure Legend Snippet: Spermine inhibits V. cholerae biofilms in an NspS/MbaA-dependent manner. Static cultures were incubated for 20 h at 27 °C in the LB only or with varying spermine concentrations. Biofilms were washed, disrupted, and measured at A 595 ( Spm , spermine). Shown are averages and S.D. ( error bars ) of three separate experiments with three technical replicates each. Means not followed by the same letter are statistically different at p ≤ 0.05 from the untreated samples within a given strain, as determined by one-way ANOVA using SigmaPlot version 12.5 ( NS , not significant).

Techniques Used: Incubation

9) Product Images from "Probiotics Streptococcus salivarius 24SMB and Streptococcus oralis 89a interfere with biofilm formation of pathogens of the upper respiratory tract"

Article Title: Probiotics Streptococcus salivarius 24SMB and Streptococcus oralis 89a interfere with biofilm formation of pathogens of the upper respiratory tract

Journal: BMC Infectious Diseases

doi: 10.1186/s12879-018-3576-9

Representative images of S. epidermidis , S. aureus and S. pneumoniae biofilms obtained by CSLM. Panels A, C and E show control biofilms, while panels B, D and F show biofilms co-cultured in presence of the probiotic strains by means of transwell inserts. Green = live cells; red = dead cells; 40× magnification
Figure Legend Snippet: Representative images of S. epidermidis , S. aureus and S. pneumoniae biofilms obtained by CSLM. Panels A, C and E show control biofilms, while panels B, D and F show biofilms co-cultured in presence of the probiotic strains by means of transwell inserts. Green = live cells; red = dead cells; 40× magnification

Techniques Used: Cell Culture

Representative images of S. pyogenes , M. catarrhalis and P. acnes biofilms obtained by CLSM. Panels A, C and E show control biofilms, while panels B, D and F show biofilms co-cultured in presence of the probiotic strains by means of transwell inserts. Green = live cells; red = dead cells; 40× magnification
Figure Legend Snippet: Representative images of S. pyogenes , M. catarrhalis and P. acnes biofilms obtained by CLSM. Panels A, C and E show control biofilms, while panels B, D and F show biofilms co-cultured in presence of the probiotic strains by means of transwell inserts. Green = live cells; red = dead cells; 40× magnification

Techniques Used: Confocal Laser Scanning Microscopy, Cell Culture

Inhibition of biofilm formation by cell-free extracts. Data are expressed as mean percentage in respect to biofilm growth control in fresh BHI broth. Black bars = control; grey tone bars = untreated cell-free extract (NT); slanted lines bars = pH neutralized cell-free extract (pH); dotted bars = heat inactivated cell-free extract (TH). * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001
Figure Legend Snippet: Inhibition of biofilm formation by cell-free extracts. Data are expressed as mean percentage in respect to biofilm growth control in fresh BHI broth. Black bars = control; grey tone bars = untreated cell-free extract (NT); slanted lines bars = pH neutralized cell-free extract (pH); dotted bars = heat inactivated cell-free extract (TH). * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Techniques Used: Inhibition

Interaction between S. salivarius 24SMB and S. oralis 89a. Data are expressed as mean absorbance ± standard deviation ( n = 3). Sal = S. salivarius; Sor = S. oralis; ** P ≤ 0.01; *** P ≤ 0.001. Panel A shows the effect of S. oralis 89a on S. salivarius 24SMB biofilm in indirect (transwell) and direct (mixed) contact. Panel B shows the effect of S. salivarius 24SMB on S. oralis 89a biofilm in indirect (transwell) and direct (mixed) contact
Figure Legend Snippet: Interaction between S. salivarius 24SMB and S. oralis 89a. Data are expressed as mean absorbance ± standard deviation ( n = 3). Sal = S. salivarius; Sor = S. oralis; ** P ≤ 0.01; *** P ≤ 0.001. Panel A shows the effect of S. oralis 89a on S. salivarius 24SMB biofilm in indirect (transwell) and direct (mixed) contact. Panel B shows the effect of S. salivarius 24SMB on S. oralis 89a biofilm in indirect (transwell) and direct (mixed) contact

Techniques Used: Standard Deviation

Inhibition of biofilm formation during time. Data are expressed as mean absorbance ± standard deviation (n = 3). Black bars = controls; grey bars = mixed co-cultures; white bars = transwell co-cultures; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001
Figure Legend Snippet: Inhibition of biofilm formation during time. Data are expressed as mean absorbance ± standard deviation (n = 3). Black bars = controls; grey bars = mixed co-cultures; white bars = transwell co-cultures; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Techniques Used: Inhibition, Standard Deviation

Inhibition of pre-formed biofilm during time. Data are expressed as mean percentage in respect to the pre-treatment level ± standard deviation (n = 3). Black bars = pre-treatment level; grey bars = 24 h; white bars = 48 h; dashed bars = 72 h; ctrl = untreated; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001
Figure Legend Snippet: Inhibition of pre-formed biofilm during time. Data are expressed as mean percentage in respect to the pre-treatment level ± standard deviation (n = 3). Black bars = pre-treatment level; grey bars = 24 h; white bars = 48 h; dashed bars = 72 h; ctrl = untreated; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Techniques Used: Inhibition, Standard Deviation

10) Product Images from "Core-satellite populations and seasonality of water meter biofilms in a metropolitan drinking water distribution system"

Article Title: Core-satellite populations and seasonality of water meter biofilms in a metropolitan drinking water distribution system

Journal: The ISME Journal

doi: 10.1038/ismej.2015.136

Diversity in biofilm (i.e., WM biofilm) and suspended (i.e., water) communities. ( a and b ) The rank-abundance distributions (close symbols: biofilm; open symbols: suspension). Dominance of high abundance and high occupancy samples are observed in both categories of samples. ( c and d ) Compare the Chao 1 ( c ) and Simpson's indices ( d ) between biofilm and suspended communities. Biofilms have lower mean values for both indices ( P -values from one-sided Welch's t- test are provided on the plot). In box plots, box represents 25 to 75 percentiles and ‘x' represents 1 and 99 percentiles.
Figure Legend Snippet: Diversity in biofilm (i.e., WM biofilm) and suspended (i.e., water) communities. ( a and b ) The rank-abundance distributions (close symbols: biofilm; open symbols: suspension). Dominance of high abundance and high occupancy samples are observed in both categories of samples. ( c and d ) Compare the Chao 1 ( c ) and Simpson's indices ( d ) between biofilm and suspended communities. Biofilms have lower mean values for both indices ( P -values from one-sided Welch's t- test are provided on the plot). In box plots, box represents 25 to 75 percentiles and ‘x' represents 1 and 99 percentiles.

Techniques Used:

Definition of core populations. Positive correlation of OTU relative abundance and occupancy in biofilm ( a ) and suspended communities ( b ) supports the use of core-satellite model. Operational definition of core communities given at different occupancy level resulted in different core community size ( c ). Number of core communities and their correlating reads are provided ( d ). Biofilm samples were subsampled by occupancy to compare with suspended communities. Average local abundance of shared (red), biofilm-only (green) and suspension-only (blue) core communities in contrast to satellite populations (gray) are provided in ( e ).
Figure Legend Snippet: Definition of core populations. Positive correlation of OTU relative abundance and occupancy in biofilm ( a ) and suspended communities ( b ) supports the use of core-satellite model. Operational definition of core communities given at different occupancy level resulted in different core community size ( c ). Number of core communities and their correlating reads are provided ( d ). Biofilm samples were subsampled by occupancy to compare with suspended communities. Average local abundance of shared (red), biofilm-only (green) and suspension-only (blue) core communities in contrast to satellite populations (gray) are provided in ( e ).

Techniques Used:

Seasonality of biofilm community structure. ( a ) Non-metrical multidimensional scaling for centroids of 20-day sampling windows. Gray eclipses indicate clusters at 40% Bray–Curtis distance. The cluster analysis result is shown as an insert. ( b ). The OTUs strongly correlated to the first CAP axis (Spearman's correlation r > 0.4 or
Figure Legend Snippet: Seasonality of biofilm community structure. ( a ) Non-metrical multidimensional scaling for centroids of 20-day sampling windows. Gray eclipses indicate clusters at 40% Bray–Curtis distance. The cluster analysis result is shown as an insert. ( b ). The OTUs strongly correlated to the first CAP axis (Spearman's correlation r > 0.4 or

Techniques Used: Sampling

11) Product Images from "Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium"

Article Title: Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00602-19

Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  
Figure Legend Snippet: Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  

Techniques Used: Activity Assay, Fluorescence

Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  
Figure Legend Snippet: Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  

Techniques Used: Activity Assay, Fluorescence, Incubation

Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.
Figure Legend Snippet: Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.

Techniques Used:

12) Product Images from "Lcl of Legionella pneumophila Is an Immunogenic GAG Binding Adhesin That Promotes Interactions with Lung Epithelial Cells and Plays a Crucial Role in Biofilm Formation ▿"

Article Title: Lcl of Legionella pneumophila Is an Immunogenic GAG Binding Adhesin That Promotes Interactions with Lung Epithelial Cells and Plays a Crucial Role in Biofilm Formation ▿

Journal: Infection and Immunity

doi: 10.1128/IAI.01304-10

Lcl promotes biofilm formation. (A) Production of biofilm by L. pneumophila Lp02 in the presence of mannose (triangles), glucose (circles), chondroitin sulfate C (squares), and fucoidan (diamonds) after 2 days of incubation. (B and C) L. pneumophila strains
Figure Legend Snippet: Lcl promotes biofilm formation. (A) Production of biofilm by L. pneumophila Lp02 in the presence of mannose (triangles), glucose (circles), chondroitin sulfate C (squares), and fucoidan (diamonds) after 2 days of incubation. (B and C) L. pneumophila strains

Techniques Used: Incubation

Confocal scanning laser microscopy (CSLM) micrographs (A) and average thicknesses (B) of biofilms produced by GFP-expressing Lp02 and isogenic Δ lpg2644 strains. A representative CSLM image is shown for each sample. Error bars represent the standard
Figure Legend Snippet: Confocal scanning laser microscopy (CSLM) micrographs (A) and average thicknesses (B) of biofilms produced by GFP-expressing Lp02 and isogenic Δ lpg2644 strains. A representative CSLM image is shown for each sample. Error bars represent the standard

Techniques Used: Microscopy, Produced, Expressing

13) Product Images from "Evaluating the Metal Tolerance Capacity of Microbial Communities Isolated from Alberta Oil Sands Process Water"

Article Title: Evaluating the Metal Tolerance Capacity of Microbial Communities Isolated from Alberta Oil Sands Process Water

Journal: PLoS ONE

doi: 10.1371/journal.pone.0148682

Heat map analysis of relative metal toxicity to (a) the OSPW consortia and (B) C . metallidurans . The heat map colors represent average minimum inhibitory concentrations (MIC) and minimum biofilm inhibitory concentrations (MBIC) based on average values obtained from two to nine trials, where red reflects the most toxic metals and green represents the least toxic. The Hard Soft Acid Base (HSAB) designation describes the behaviour of metal ions based on preferential donor ligands. Soft acids prefer to bind with thiol (S-group) ligands, hard acids with N and O, and borderline acids have varied preference for S, N, and O-containing ligands.
Figure Legend Snippet: Heat map analysis of relative metal toxicity to (a) the OSPW consortia and (B) C . metallidurans . The heat map colors represent average minimum inhibitory concentrations (MIC) and minimum biofilm inhibitory concentrations (MBIC) based on average values obtained from two to nine trials, where red reflects the most toxic metals and green represents the least toxic. The Hard Soft Acid Base (HSAB) designation describes the behaviour of metal ions based on preferential donor ligands. Soft acids prefer to bind with thiol (S-group) ligands, hard acids with N and O, and borderline acids have varied preference for S, N, and O-containing ligands.

Techniques Used:

14) Product Images from "Spermine inhibits Vibrio cholerae biofilm formation through the NspS–MbaA polyamine signaling system"

Article Title: Spermine inhibits Vibrio cholerae biofilm formation through the NspS–MbaA polyamine signaling system

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M117.801068

Spermine inhibits vpsL gene transcription in a NspS—MbaA-dependent manner. Static cultures were grown for 20 h with the indicated concentration of spermine ( Spm ) at 27 °C. Planktonic cells and biofilms were homogenized by disruption with glass beads and vortexing. Cells were pelleted and lysed in Z-buffer. Lysates were incubated with the colorimetric β-galactosidase substrate ortho -nitrophenyl-β-galactoside, and Miller units were calculated using A 415 of reaction mixtures. Shown are the averages and S.D. ( error bars ) of three experiments with three technical replicates each. Means not followed by the same letter are statistically different at p ≤ 0.05 from the untreated samples within a given strain, as determined by one-way ANOVA using SigmaPlot version 12.5 ( NS , not significant).
Figure Legend Snippet: Spermine inhibits vpsL gene transcription in a NspS—MbaA-dependent manner. Static cultures were grown for 20 h with the indicated concentration of spermine ( Spm ) at 27 °C. Planktonic cells and biofilms were homogenized by disruption with glass beads and vortexing. Cells were pelleted and lysed in Z-buffer. Lysates were incubated with the colorimetric β-galactosidase substrate ortho -nitrophenyl-β-galactoside, and Miller units were calculated using A 415 of reaction mixtures. Shown are the averages and S.D. ( error bars ) of three experiments with three technical replicates each. Means not followed by the same letter are statistically different at p ≤ 0.05 from the untreated samples within a given strain, as determined by one-way ANOVA using SigmaPlot version 12.5 ( NS , not significant).

Techniques Used: Concentration Assay, Incubation

Spermine inhibits V. cholerae biofilms in an NspS/MbaA-dependent manner. Static cultures were incubated for 20 h at 27 °C in the LB only or with varying spermine concentrations. Biofilms were washed, disrupted, and measured at A 595 ( Spm , spermine). Shown are averages and S.D. ( error bars ) of three separate experiments with three technical replicates each. Means not followed by the same letter are statistically different at p ≤ 0.05 from the untreated samples within a given strain, as determined by one-way ANOVA using SigmaPlot version 12.5 ( NS , not significant).
Figure Legend Snippet: Spermine inhibits V. cholerae biofilms in an NspS/MbaA-dependent manner. Static cultures were incubated for 20 h at 27 °C in the LB only or with varying spermine concentrations. Biofilms were washed, disrupted, and measured at A 595 ( Spm , spermine). Shown are averages and S.D. ( error bars ) of three separate experiments with three technical replicates each. Means not followed by the same letter are statistically different at p ≤ 0.05 from the untreated samples within a given strain, as determined by one-way ANOVA using SigmaPlot version 12.5 ( NS , not significant).

Techniques Used: Incubation

15) Product Images from "Comparison of the Antibiotic Activities of Daptomycin, Vancomycin, and the Investigational Fluoroquinolone Delafloxacin against Biofilms from Staphylococcus aureus Clinical Isolates"

Article Title: Comparison of the Antibiotic Activities of Daptomycin, Vancomycin, and the Investigational Fluoroquinolone Delafloxacin against Biofilms from Staphylococcus aureus Clinical Isolates

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.03482-14

Penetration of antibiotics within biofilms. Confocal images of biofilms incubated for 1 h with 50 mg/liter delafloxacin (top [blue]), 20 mg/liter Bodipy-FL-daptomycin (middle [green]), or 20 mg/liter Bodipy-FL-vancomycin (bottom [green]) and labeled with LIVE/DEAD stain (top: red, dead; green, live) or CTC (middle and bottom: red). The graphs below each column compare the relative penetration of the drugs within the depth of the corresponding biofilm, expressed as the percentage of the added concentration (DFX, delafloxacin; DAP, daptomycin; VAN, vancomycin).
Figure Legend Snippet: Penetration of antibiotics within biofilms. Confocal images of biofilms incubated for 1 h with 50 mg/liter delafloxacin (top [blue]), 20 mg/liter Bodipy-FL-daptomycin (middle [green]), or 20 mg/liter Bodipy-FL-vancomycin (bottom [green]) and labeled with LIVE/DEAD stain (top: red, dead; green, live) or CTC (middle and bottom: red). The graphs below each column compare the relative penetration of the drugs within the depth of the corresponding biofilm, expressed as the percentage of the added concentration (DFX, delafloxacin; DAP, daptomycin; VAN, vancomycin).

Techniques Used: Incubation, Labeling, Staining, Concentration Assay

Activities of antibiotics against biofilms. Concentration-response activities of antibiotics against 24-h biofilms of strain 2011S027 (top) or 2003/651 (bottom). Twenty-four-hour biofilms were incubated with increasing concentrations of antibiotics for 48 h (DFX, delafloxacin; DAP, daptomycin; VAN, vancomycin). The ordinate shows the change in viability (assessed by resorufin fluorescence; left) or in biomass (assessed by crystal violet absorbance; right) as the percentage of the control (CT) value (no antibiotic present). All values are the means ± standard deviations (SD) of 8 wells (when not visible, the SD bars are smaller than the size of the symbols).
Figure Legend Snippet: Activities of antibiotics against biofilms. Concentration-response activities of antibiotics against 24-h biofilms of strain 2011S027 (top) or 2003/651 (bottom). Twenty-four-hour biofilms were incubated with increasing concentrations of antibiotics for 48 h (DFX, delafloxacin; DAP, daptomycin; VAN, vancomycin). The ordinate shows the change in viability (assessed by resorufin fluorescence; left) or in biomass (assessed by crystal violet absorbance; right) as the percentage of the control (CT) value (no antibiotic present). All values are the means ± standard deviations (SD) of 8 wells (when not visible, the SD bars are smaller than the size of the symbols).

Techniques Used: Concentration Assay, Incubation, Fluorescence

Biofilm characterization. Resorufin fluorescence signal (RF) (left), crystal violet absorbance (CV) (middle), and calcofluor white fluorescence (CFW) signal (right, squares) measured in 24-h biofilm. The right axis in the right graph shows CFW fluorescence values normalized with respect to biomass, as evaluated by CV absorbance (diamonds). The data are the means ± standard deviations (SD) of 8 wells. Statistical analysis was performed using an analysis of variance (ANOVA) with Dunnett's post hoc test; the strains with different letters for each data set denote statistically significant differences among them (*, P
Figure Legend Snippet: Biofilm characterization. Resorufin fluorescence signal (RF) (left), crystal violet absorbance (CV) (middle), and calcofluor white fluorescence (CFW) signal (right, squares) measured in 24-h biofilm. The right axis in the right graph shows CFW fluorescence values normalized with respect to biomass, as evaluated by CV absorbance (diamonds). The data are the means ± standard deviations (SD) of 8 wells. Statistical analysis was performed using an analysis of variance (ANOVA) with Dunnett's post hoc test; the strains with different letters for each data set denote statistically significant differences among them (*, P

Techniques Used: Fluorescence

Influence of polyamines on antibiotic activity against biofilm of strain 2003/651. Left, concentration-response activities of antibiotics against 24-h biofilms incubated with increasing concentrations of antibiotics for 48 h (DFX, delafloxacin; DAP, daptomycin; VAN, vancomycin) in the absence or presence of 200 μM norspermine or norspermidine. The ordinate shows the change in viability (assessed by resorufin fluorescence; left) or in biomass (assessed by crystal violet absorbance; right) as the percentage of the control value (no antibiotic present). All values are the means ± standard deviations (SD) of 8 wells (when not visible, the SD bars are smaller than the size of the symbols). The arrows point to the concentration of antibiotic used in confocal microscopy. Middle, confocal images of biofilms incubated for 1 h with 50 mg/liter delafloxacin (blue), 20 mg/liter Bodipy-FL-daptomycin (green), or 20 mg/liter Bodipy-FL-vancomycin (green) in the absence or presence of 200 μM norspermine and labeled with LIVE/DEAD staining (top: red, dead; green, live) or CTC (middle and bottom: red). Right, relative penetration (in mg/liter) of the drugs within the depth of the corresponding biofilms, under control conditions, or in the presence of 200 μM norspermidine. The horizontal dotted line corresponds to the MIC of each antibiotic.
Figure Legend Snippet: Influence of polyamines on antibiotic activity against biofilm of strain 2003/651. Left, concentration-response activities of antibiotics against 24-h biofilms incubated with increasing concentrations of antibiotics for 48 h (DFX, delafloxacin; DAP, daptomycin; VAN, vancomycin) in the absence or presence of 200 μM norspermine or norspermidine. The ordinate shows the change in viability (assessed by resorufin fluorescence; left) or in biomass (assessed by crystal violet absorbance; right) as the percentage of the control value (no antibiotic present). All values are the means ± standard deviations (SD) of 8 wells (when not visible, the SD bars are smaller than the size of the symbols). The arrows point to the concentration of antibiotic used in confocal microscopy. Middle, confocal images of biofilms incubated for 1 h with 50 mg/liter delafloxacin (blue), 20 mg/liter Bodipy-FL-daptomycin (green), or 20 mg/liter Bodipy-FL-vancomycin (green) in the absence or presence of 200 μM norspermine and labeled with LIVE/DEAD staining (top: red, dead; green, live) or CTC (middle and bottom: red). Right, relative penetration (in mg/liter) of the drugs within the depth of the corresponding biofilms, under control conditions, or in the presence of 200 μM norspermidine. The horizontal dotted line corresponds to the MIC of each antibiotic.

Techniques Used: Activity Assay, Concentration Assay, Incubation, Fluorescence, Confocal Microscopy, Labeling, Staining

16) Product Images from "Copper and Quaternary Ammonium Cations Exert Synergistic Bactericidal and Antibiofilm Activity against Pseudomonas aeruginosa "

Article Title: Copper and Quaternary Ammonium Cations Exert Synergistic Bactericidal and Antibiofilm Activity against Pseudomonas aeruginosa

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00203-08

Combinations of Cu 2+ with other QACs show synergistic killing of P. aeruginosa ATCC 15442 biofilms. Viable cell counts were determined after exposure of biofilms to combinations of Cu 2+ and benzalkonium chloride (a), cetylpyridinium chloride
Figure Legend Snippet: Combinations of Cu 2+ with other QACs show synergistic killing of P. aeruginosa ATCC 15442 biofilms. Viable cell counts were determined after exposure of biofilms to combinations of Cu 2+ and benzalkonium chloride (a), cetylpyridinium chloride

Techniques Used:

Live/Dead BacLight staining of P. aeruginosa ATCC 27853 biofilms exposed to Cu 2+ and Polycide indicates that these agents are bactericidal. In these pictures, green cells are live bacteria and red cells are dead bacteria. (a) Growth controls.
Figure Legend Snippet: Live/Dead BacLight staining of P. aeruginosa ATCC 27853 biofilms exposed to Cu 2+ and Polycide indicates that these agents are bactericidal. In these pictures, green cells are live bacteria and red cells are dead bacteria. (a) Growth controls.

Techniques Used: Staining

High-throughput screening may be used to identify synergistic antimicrobial interactions that kill microbial biofilms. Starting from cryogenic stocks, the desired bacterial strain was streaked out twice on TSA (a), and colonies from the second subcultures
Figure Legend Snippet: High-throughput screening may be used to identify synergistic antimicrobial interactions that kill microbial biofilms. Starting from cryogenic stocks, the desired bacterial strain was streaked out twice on TSA (a), and colonies from the second subcultures

Techniques Used: High Throughput Screening Assay

P. aeruginosa ATCC 15442 biofilms were killed time dependently by combinations of Cu 2+ and Polycide. Viable cell counts were determined after exposure of biofilms to combinations of Cu 2+ and Polycide in ddH 2 O for 10 min (a) or 30 min (b)
Figure Legend Snippet: P. aeruginosa ATCC 15442 biofilms were killed time dependently by combinations of Cu 2+ and Polycide. Viable cell counts were determined after exposure of biofilms to combinations of Cu 2+ and Polycide in ddH 2 O for 10 min (a) or 30 min (b)

Techniques Used:

Killing of Escherichia coli (a), Pseudomonas fluorescens (b), Salmonella enterica serovar Cholerasuis (c), and Staphylococcus aureus (d) biofilms by combinations of Cu 2+ and Polycide. These data are for 24 h of exposure in dilute organics, and
Figure Legend Snippet: Killing of Escherichia coli (a), Pseudomonas fluorescens (b), Salmonella enterica serovar Cholerasuis (c), and Staphylococcus aureus (d) biofilms by combinations of Cu 2+ and Polycide. These data are for 24 h of exposure in dilute organics, and

Techniques Used:

17) Product Images from "Fungal β-1,3-Glucan Increases Ofloxacin Tolerance of Escherichia coli in a Polymicrobial E. coli/ Candida albicans Biofilm"

Article Title: Fungal β-1,3-Glucan Increases Ofloxacin Tolerance of Escherichia coli in a Polymicrobial E. coli/ Candida albicans Biofilm

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.04650-14

Increased ofloxacin tolerance of E. coli in an E. coli/C. albicans biofilm. E. coli (gray bars) and E. coli/C. albicans (black bars) biofilms were treated with different concentrations of ofloxacin (0.39 to 3.13 μM). Afterwards, survival of E. coli was quantified using selective plating. *, P
Figure Legend Snippet: Increased ofloxacin tolerance of E. coli in an E. coli/C. albicans biofilm. E. coli (gray bars) and E. coli/C. albicans (black bars) biofilms were treated with different concentrations of ofloxacin (0.39 to 3.13 μM). Afterwards, survival of E. coli was quantified using selective plating. *, P

Techniques Used:

Increased ofloxacin tolerance of E. coli in the presence of C. albicans is mainly biofilm specific. Survival of E. coli in planktonic conditions upon ofloxacin treatment (0.09 to 0.75 μM) was quantified in the absence (gray bars) or presence (black bars) of C. albicans . *, P
Figure Legend Snippet: Increased ofloxacin tolerance of E. coli in the presence of C. albicans is mainly biofilm specific. Survival of E. coli in planktonic conditions upon ofloxacin treatment (0.09 to 0.75 μM) was quantified in the absence (gray bars) or presence (black bars) of C. albicans . *, P

Techniques Used:

The presence of C. albicans zap1 Δ/ zap1 Δ increases ofloxacin tolerance of E. coli to a greater extent than does the presence of C. albicans wild type. E. coli/C. albicans biofilms consisting of E. coli wild type and C. albicans wild type (black bars) or zap1 Δ/ zap1 Δ deletion mutant (gray bars) were treated with 0.78 μM ofloxacin. Afterwards, survival of E. coli was quantified using selective plating. Statistical analysis was performed using an unpaired t test. ***, P
Figure Legend Snippet: The presence of C. albicans zap1 Δ/ zap1 Δ increases ofloxacin tolerance of E. coli to a greater extent than does the presence of C. albicans wild type. E. coli/C. albicans biofilms consisting of E. coli wild type and C. albicans wild type (black bars) or zap1 Δ/ zap1 Δ deletion mutant (gray bars) were treated with 0.78 μM ofloxacin. Afterwards, survival of E. coli was quantified using selective plating. Statistical analysis was performed using an unpaired t test. ***, P

Techniques Used: Mutagenesis

The extracellular matrix contributes to the observed increased ofloxacin tolerance of E. coli in an E. coli/C. albicans biofilm. E. coli/C. albicans and E. coli biofilms were treated with 0.78 μM ofloxacin with or without matrix-degrading enzymes (50 μg/ml). Afterwards, survival of E. coli was quantified using selective plating. *, P
Figure Legend Snippet: The extracellular matrix contributes to the observed increased ofloxacin tolerance of E. coli in an E. coli/C. albicans biofilm. E. coli/C. albicans and E. coli biofilms were treated with 0.78 μM ofloxacin with or without matrix-degrading enzymes (50 μg/ml). Afterwards, survival of E. coli was quantified using selective plating. *, P

Techniques Used:

Interaction of C. albicans and E. coli in E. coli/C. albicans biofilms. E. coli and C. albicans were grown for 24 h at 37°C using titanium disks as the substrate. After dehydration, samples were visualized using SEM.
Figure Legend Snippet: Interaction of C. albicans and E. coli in E. coli/C. albicans biofilms. E. coli and C. albicans were grown for 24 h at 37°C using titanium disks as the substrate. After dehydration, samples were visualized using SEM.

Techniques Used:

Exogenously added laminarin increases ofloxacin tolerance of E. coli in an E. coli biofilm. An E. coli biofilm was treated with different concentrations of ofloxacin in the presence or absence of different concentrations of laminarin (0 to 0.5 mg/ml). Biomass was quantified using crystal violet. **, P
Figure Legend Snippet: Exogenously added laminarin increases ofloxacin tolerance of E. coli in an E. coli biofilm. An E. coli biofilm was treated with different concentrations of ofloxacin in the presence or absence of different concentrations of laminarin (0 to 0.5 mg/ml). Biomass was quantified using crystal violet. **, P

Techniques Used:

18) Product Images from "Activities of Combinations of Antistaphylococcal Antibiotics with Fusidic Acid against Staphylococcal Biofilms in In Vitro Static and Dynamic Models"

Article Title: Activities of Combinations of Antistaphylococcal Antibiotics with Fusidic Acid against Staphylococcal Biofilms in In Vitro Static and Dynamic Models

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00598-18

Activity of fusidic acid (FUS) alone or combined with selected antibiotics (daptomycin [DAP], linezolid [LZD], vancomycin [VAN], rifampin [RIF], moxifloxacin [MXF], and doxycycline [DOX]) after 48 h of incubation with biofilms from the clinical isolate 80224422456. The graphs show the residual viability evaluated by the reduction in resorufin fluorescence compared to that of the untreated control (CT) as a function of the concentration of fusidic acid. Combined antibiotics were added at a fixed concentration corresponding to their human fC max . The horizontal, colored dotted line in each panel shows the effect of the combined antibiotic alone at the selected concentration; therefore, all data points located in the colored area correspond to activities higher than that of the combined antibiotic alone. The black vertical dotted lines show the fC min and fC max of fusidic acid. All data are the means ± SDs from quadruplicates.
Figure Legend Snippet: Activity of fusidic acid (FUS) alone or combined with selected antibiotics (daptomycin [DAP], linezolid [LZD], vancomycin [VAN], rifampin [RIF], moxifloxacin [MXF], and doxycycline [DOX]) after 48 h of incubation with biofilms from the clinical isolate 80224422456. The graphs show the residual viability evaluated by the reduction in resorufin fluorescence compared to that of the untreated control (CT) as a function of the concentration of fusidic acid. Combined antibiotics were added at a fixed concentration corresponding to their human fC max . The horizontal, colored dotted line in each panel shows the effect of the combined antibiotic alone at the selected concentration; therefore, all data points located in the colored area correspond to activities higher than that of the combined antibiotic alone. The black vertical dotted lines show the fC min and fC max of fusidic acid. All data are the means ± SDs from quadruplicates.

Techniques Used: Activity Assay, Incubation, Fluorescence, Concentration Assay

Activity of fusidic acid (FUS) alone or combined with daptomycin (DAP), linezolid (LZD), or vancomycin (VAN) against biofilms from the reference strain ATCC 25923 (top) or the clinical isolate 80224422456 (bottom) in the CDC bioreactor. FUS, LZD, and VAN were injected twice (0 h and 12 h), and DAP once (0 h), at the following concentrations (to mimic fC max ): FUS, 14 mg/liter; DAP, 9.8 mg/liter; LZD, 17 mg/liter; VAN, 20 mg/liter. Peristaltic pumps were then started to infuse antibiotic-free medium at rate set up to mimic a t 1/2 of 12 h for FUS, 8 h for DAP, and 6 h for LZD and VAN.
Figure Legend Snippet: Activity of fusidic acid (FUS) alone or combined with daptomycin (DAP), linezolid (LZD), or vancomycin (VAN) against biofilms from the reference strain ATCC 25923 (top) or the clinical isolate 80224422456 (bottom) in the CDC bioreactor. FUS, LZD, and VAN were injected twice (0 h and 12 h), and DAP once (0 h), at the following concentrations (to mimic fC max ): FUS, 14 mg/liter; DAP, 9.8 mg/liter; LZD, 17 mg/liter; VAN, 20 mg/liter. Peristaltic pumps were then started to infuse antibiotic-free medium at rate set up to mimic a t 1/2 of 12 h for FUS, 8 h for DAP, and 6 h for LZD and VAN.

Techniques Used: Activity Assay, Injection

Activity of fusidic acid (FUS) alone or combined with selected antibiotics (daptomycin [DAP], linezolid [LZD], and vancomycin [VAN]) after 48 h of incubation with biofilms from ATCC 25923. The graphs show the residual viability evaluated by the reduction in resorufin fluorescence compared to that of the untreated control (CT) as a function of the concentration of fusidic acid. Combined antibiotics were added at a fixed concentration corresponding to their human fC min (top) or fC max (bottom). The horizontal, colored dotted line in each panel shows the effect of the combined antibiotic used alone at the selected concentration; therefore, all data points located in the colored area correspond to activities higher than that measured for this combined antibiotic alone. The black vertical dotted lines show the fC min and fC max of fusidic acid. All data are the means ± SDs from quadruplicates.
Figure Legend Snippet: Activity of fusidic acid (FUS) alone or combined with selected antibiotics (daptomycin [DAP], linezolid [LZD], and vancomycin [VAN]) after 48 h of incubation with biofilms from ATCC 25923. The graphs show the residual viability evaluated by the reduction in resorufin fluorescence compared to that of the untreated control (CT) as a function of the concentration of fusidic acid. Combined antibiotics were added at a fixed concentration corresponding to their human fC min (top) or fC max (bottom). The horizontal, colored dotted line in each panel shows the effect of the combined antibiotic used alone at the selected concentration; therefore, all data points located in the colored area correspond to activities higher than that measured for this combined antibiotic alone. The black vertical dotted lines show the fC min and fC max of fusidic acid. All data are the means ± SDs from quadruplicates.

Techniques Used: Activity Assay, Incubation, Fluorescence, Concentration Assay

19) Product Images from "Bacteriophage Lysin CF-301, a Potent Antistaphylococcal Biofilm Agent"

Article Title: Bacteriophage Lysin CF-301, a Potent Antistaphylococcal Biofilm Agent

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.02666-16

Microscopic analysis of 24-h-old biofilms on glass surfaces. (A) Confocal analysis of GFP-expressing MSSA strain Newman (CFS-1246) treated for 1 h with buffer or CF-301 (32 μg/ml). The box size is 290 μm by 290 μm by 25 μm, and voxel dimensions are 0.284 μm by 0.284 μm by 0.420 μm (scale bars = 40.0 μm). (B) Biofilms of ATCC BAA-42 on glass chamber slides, treated for 1 h with buffer or CF-301 (32 μg/ml), and stained with FilmTracer Calcein green biofilm stain. DIC images are shown with and without fluorescence (magnification, ×400). (C) Unmagnified biofilms of MRSA strain ATCC BAA-42 on glass chamber slides, treated for 1 h with buffer or CF-301 (32 μg/ml). After treatment, biofilm CFU were determined.
Figure Legend Snippet: Microscopic analysis of 24-h-old biofilms on glass surfaces. (A) Confocal analysis of GFP-expressing MSSA strain Newman (CFS-1246) treated for 1 h with buffer or CF-301 (32 μg/ml). The box size is 290 μm by 290 μm by 25 μm, and voxel dimensions are 0.284 μm by 0.284 μm by 0.420 μm (scale bars = 40.0 μm). (B) Biofilms of ATCC BAA-42 on glass chamber slides, treated for 1 h with buffer or CF-301 (32 μg/ml), and stained with FilmTracer Calcein green biofilm stain. DIC images are shown with and without fluorescence (magnification, ×400). (C) Unmagnified biofilms of MRSA strain ATCC BAA-42 on glass chamber slides, treated for 1 h with buffer or CF-301 (32 μg/ml). After treatment, biofilm CFU were determined.

Techniques Used: Expressing, Staining, Fluorescence

Disruption of catheter biofilms. Biofilms of MRSA strain ATCC BAA-42 were grown for 3 days in the catheter lumen (covering half of the internal diameter) before treatment for 4 h with buffer, daptomycin (DAP; 1 μg/ml and 1 mg/ml), or CF-301 (32 μg/ml). (A) Methylene blue-stained biofilm biomass remaining after treatment. (B) Representation of catheter biofilms, cut longitudinally to expose the internal biomass. Top-down and leading-edge views are indicated. (C) Top-down view of methylene blue-stained biofilm biomass. Bright-field microscopic images are shown at ×400 magnification. (D) Leading-edge view of methylene blue-stained biofilms. Darkly stained material represents the biofilm. Bright-field microscopic images are shown at ×400 magnification. (E) SEM images of catheter biofilms remaining after the indicated treatments. The following magnifications were used: buffer, ×500; DAP, ×7,500; CF-301, ×7,500; and no biofilm, ×7,500.
Figure Legend Snippet: Disruption of catheter biofilms. Biofilms of MRSA strain ATCC BAA-42 were grown for 3 days in the catheter lumen (covering half of the internal diameter) before treatment for 4 h with buffer, daptomycin (DAP; 1 μg/ml and 1 mg/ml), or CF-301 (32 μg/ml). (A) Methylene blue-stained biofilm biomass remaining after treatment. (B) Representation of catheter biofilms, cut longitudinally to expose the internal biomass. Top-down and leading-edge views are indicated. (C) Top-down view of methylene blue-stained biofilm biomass. Bright-field microscopic images are shown at ×400 magnification. (D) Leading-edge view of methylene blue-stained biofilms. Darkly stained material represents the biofilm. Bright-field microscopic images are shown at ×400 magnification. (E) SEM images of catheter biofilms remaining after the indicated treatments. The following magnifications were used: buffer, ×500; DAP, ×7,500; CF-301, ×7,500; and no biofilm, ×7,500.

Techniques Used: Staining

SEM images of catheter biofilms treated with CF-301. Biofilms of MRSA strain ATCC BAA-42 were grown for 3 days in the catheter lumen prior to either a 30-s or 15-min treatment with CF-301 (32 μg/ml). A 15-min buffer treatment was included as a control, and all magnifications are indicated.
Figure Legend Snippet: SEM images of catheter biofilms treated with CF-301. Biofilms of MRSA strain ATCC BAA-42 were grown for 3 days in the catheter lumen prior to either a 30-s or 15-min treatment with CF-301 (32 μg/ml). A 15-min buffer treatment was included as a control, and all magnifications are indicated.

Techniques Used:

Titration and time course analysis of CF-301-mediated biofilm biomass disruption and associated CFU reduction. Biofilms of MRSA strain ATCC BAA-42 were grown for 3 days in the catheter lumen before treatment. (A) Biofilms treated for 4 h with buffer, DAP, or CF-301. Drug concentrations are based on MIC values; MICs of DAP and CF-301 are 1 μg/ml and 32 μg/ml, respectively. Methylene blue-stained biofilms are shown with a matched analysis of biofilm CFU remaining after each treatment. Asterisks denote the limit of detection. (B) Biofilms treated with CF-301 (32 μg/ml) over a 4-h time course. At the indicated time points, catheters were washed and biofilms were either stained with methylene blue or quantitated to determine the CFU. Asterisks denote the limit of detection.
Figure Legend Snippet: Titration and time course analysis of CF-301-mediated biofilm biomass disruption and associated CFU reduction. Biofilms of MRSA strain ATCC BAA-42 were grown for 3 days in the catheter lumen before treatment. (A) Biofilms treated for 4 h with buffer, DAP, or CF-301. Drug concentrations are based on MIC values; MICs of DAP and CF-301 are 1 μg/ml and 32 μg/ml, respectively. Methylene blue-stained biofilms are shown with a matched analysis of biofilm CFU remaining after each treatment. Asterisks denote the limit of detection. (B) Biofilms treated with CF-301 (32 μg/ml) over a 4-h time course. At the indicated time points, catheters were washed and biofilms were either stained with methylene blue or quantitated to determine the CFU. Asterisks denote the limit of detection.

Techniques Used: Titration, Staining

Disruption of S. aureus biofilm biomass. Biofilms of MRSA strain ATCC BAA-42 were grown over 24 h (A to C) or 2 weeks (D) in 24-well polystyrene plates and treated with either CF-301 (MIC = 32 μg/ml) or antibiotics (1,000× the MIC = 1 mg/ml) in MHB or CA-MHB. Medium-alone controls were included. (A) Crystal violet staining of biofilms treated for 24 h. (B) Quantitation of crystal violet staining over a 24-h time course. Assays were performed in triplicate, and OD 600 data are the mean ± standard deviation. (C) Biofilms treated with a 2-fold dilution series of CF-301 for 4 h prior to crystal violet staining and quantitation. (D) Biofilms treated with a 10-fold dilution series of CF-301 or DAP for 4 or 24 h prior to staining with crystal violet. LZD, linezolid.
Figure Legend Snippet: Disruption of S. aureus biofilm biomass. Biofilms of MRSA strain ATCC BAA-42 were grown over 24 h (A to C) or 2 weeks (D) in 24-well polystyrene plates and treated with either CF-301 (MIC = 32 μg/ml) or antibiotics (1,000× the MIC = 1 mg/ml) in MHB or CA-MHB. Medium-alone controls were included. (A) Crystal violet staining of biofilms treated for 24 h. (B) Quantitation of crystal violet staining over a 24-h time course. Assays were performed in triplicate, and OD 600 data are the mean ± standard deviation. (C) Biofilms treated with a 2-fold dilution series of CF-301 for 4 h prior to crystal violet staining and quantitation. (D) Biofilms treated with a 10-fold dilution series of CF-301 or DAP for 4 or 24 h prior to staining with crystal violet. LZD, linezolid.

Techniques Used: Staining, Quantitation Assay, Standard Deviation

Disruption of S. aureus biofilms enriched for SCVs. To select for SCVs, biofilms of MRSA strain BAA-42 were formed on 24-well plates over 3 days in the presence of daptomycin (DAP; 1 μg/ml), vancomycin (VAN; 1 μg/ml), or oxacillin (OXA; 0.5 μg/ml). (A) Appearance of SCVs among bacteria recovered from biofilms grown with and without DAP. SCVs are indicated by arrows. (B) Crystal violet staining of biofilms after growth in the presence and absence of DAP and treatment with CF-301 for 24 h. The results of duplicate experiments are shown. (C) Quantitation of crystal violet staining of biofilms grown in the presence and absence of DAP and treated with CF-301 for 24 h. Values were determined as a percentage of the buffer-treated (i.e., no CF-301) control. (D) Quantitation of crystal violet staining of biofilms grown in the presence and absence of VAN or OXA and treated with a CF-301 for 24 h. Values were determined as a percentage of the buffer-treated (i.e., no CF-301) control. (E) CF-301-mediated killing of biofilm bacteria grown in the presence and absence of DAP. After 24-h treatment with CF-301, both the biofilm and planktonic cells were quantitated. Asterisks denote the limit of detection.
Figure Legend Snippet: Disruption of S. aureus biofilms enriched for SCVs. To select for SCVs, biofilms of MRSA strain BAA-42 were formed on 24-well plates over 3 days in the presence of daptomycin (DAP; 1 μg/ml), vancomycin (VAN; 1 μg/ml), or oxacillin (OXA; 0.5 μg/ml). (A) Appearance of SCVs among bacteria recovered from biofilms grown with and without DAP. SCVs are indicated by arrows. (B) Crystal violet staining of biofilms after growth in the presence and absence of DAP and treatment with CF-301 for 24 h. The results of duplicate experiments are shown. (C) Quantitation of crystal violet staining of biofilms grown in the presence and absence of DAP and treated with CF-301 for 24 h. Values were determined as a percentage of the buffer-treated (i.e., no CF-301) control. (D) Quantitation of crystal violet staining of biofilms grown in the presence and absence of VAN or OXA and treated with a CF-301 for 24 h. Values were determined as a percentage of the buffer-treated (i.e., no CF-301) control. (E) CF-301-mediated killing of biofilm bacteria grown in the presence and absence of DAP. After 24-h treatment with CF-301, both the biofilm and planktonic cells were quantitated. Asterisks denote the limit of detection.

Techniques Used: Staining, Quantitation Assay

Formation of mixed-species biofilms and analysis of CF-301 susceptibility. Biofilms containing S. aureus (strain BAA-42) and/or S. epidermidis (strain NRS 7) were grown either on 24-well polystyrene plates for 3 days or on catheters and surgical mesh for 6 days before treatment with a 10-fold dilution series of CF-301. (A) Quantitation of S. aureus and S. epidermidis CFU in mixed biofilms prior to CF-301 treatment. Experiments were performed in triplicate, and CFU represent the mean ± standard deviation. (B) Crystal violet staining of single- and mixed-species biofilms on 24-well plates treated for 24 h with CF-301 (relative to untreated control). (C) Catheter biofilms treated for 4 h. (D) Catheter biofilms treated for 24 h. (E) Surgical mesh biofilms treated for 24 h.
Figure Legend Snippet: Formation of mixed-species biofilms and analysis of CF-301 susceptibility. Biofilms containing S. aureus (strain BAA-42) and/or S. epidermidis (strain NRS 7) were grown either on 24-well polystyrene plates for 3 days or on catheters and surgical mesh for 6 days before treatment with a 10-fold dilution series of CF-301. (A) Quantitation of S. aureus and S. epidermidis CFU in mixed biofilms prior to CF-301 treatment. Experiments were performed in triplicate, and CFU represent the mean ± standard deviation. (B) Crystal violet staining of single- and mixed-species biofilms on 24-well plates treated for 24 h with CF-301 (relative to untreated control). (C) Catheter biofilms treated for 4 h. (D) Catheter biofilms treated for 24 h. (E) Surgical mesh biofilms treated for 24 h.

Techniques Used: Quantitation Assay, Standard Deviation, Staining

20) Product Images from "Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium"

Article Title: Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00602-19

Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  
Figure Legend Snippet: Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  

Techniques Used: Activity Assay, Fluorescence

Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  
Figure Legend Snippet: Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P  

Techniques Used: Activity Assay, Fluorescence, Incubation

Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.
Figure Legend Snippet: Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.

Techniques Used:

21) Product Images from "Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium"

Article Title: Activity of Antibiotics against Staphylococcus aureus in an In Vitro Model of Biofilms in the Context of Cystic Fibrosis: Influence of the Culture Medium

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00602-19

Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P
Figure Legend Snippet: Comparison of antibiotic maximal efficacies ( E max ) expressed as the reduction in the number of CFU from that for the control (left) or as a percentage of the reduction in metabolic activity (resorufin fluorescence; middle) or biofilm mass (crystal violet [CV] absorbance; right) compared to that for an untreated biofilm for strain ATCC 25923 (open bar) or ATCC 33591 (closed bars) grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). MEM, meropenem; VAN, vancomycin; LZD, linezolid; AZM, azithromycin; RIF, rifampin; CIP, ciprofloxacin; TOB, tobramycin. Values are means ± SEM. Statistical analyses were by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P

Techniques Used: Activity Assay, Fluorescence

Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P
Figure Legend Snippet: Metabolic activity in planktonic cultures or biofilm cultures grown in Trypticase soy broth supplemented with 1% glucose and 2% NaCl (TGN) or in artificial sputum medium (ASM). (Left) Resorufin fluorescence signal recorded after 30 min of incubation of planktonic bacteria at increasing inocula with 10-mg/liter resazurin. (Right) Resorufin fluorescence signal recorded after 30 min of incubation of a 24-h-old biofilm with 10-mg/liter resazurin. Data are means ± SD for triplicates in a single experiment or means ± SEM from at least 3 independent experiments performed in triplicate. Statistical analyses comparing strains in each individual medium were performed by one-way analysis of variance (ANOVA) with Tukey’s posttest for multiple comparisons; values with different letters are significantly different from each other ( P

Techniques Used: Activity Assay, Fluorescence, Incubation

Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.
Figure Legend Snippet: Morphology and counts of colonies from biofilms of ATCC 25923 cultivated in artificial sputum medium (ASM) and exposed for 24 h to tobramycin (TOB) at 5 (TOB 5), 10 (TOB 10), or 100 (TOB 100) mg/liter or under control conditions (no antibiotic added as a control [CT]). Samples were plated on Columbia blood agar, TSA, or TSA supplemented with either 1 mg/liter hemin (TSAH), menadione (TSAM). or both hemin and menadione (TSAMH). Yellow arrows indicate a typical small colony.

Techniques Used:

22) Product Images from "Probiotics Streptococcus salivarius 24SMB and Streptococcus oralis 89a interfere with biofilm formation of pathogens of the upper respiratory tract"

Article Title: Probiotics Streptococcus salivarius 24SMB and Streptococcus oralis 89a interfere with biofilm formation of pathogens of the upper respiratory tract

Journal: BMC Infectious Diseases

doi: 10.1186/s12879-018-3576-9

Representative images of S. epidermidis , S. aureus and S. pneumoniae biofilms obtained by CSLM. Panels A, C and E show control biofilms, while panels B, D and F show biofilms co-cultured in presence of the probiotic strains by means of transwell inserts. Green = live cells; red = dead cells; 40× magnification
Figure Legend Snippet: Representative images of S. epidermidis , S. aureus and S. pneumoniae biofilms obtained by CSLM. Panels A, C and E show control biofilms, while panels B, D and F show biofilms co-cultured in presence of the probiotic strains by means of transwell inserts. Green = live cells; red = dead cells; 40× magnification

Techniques Used: Cell Culture

Representative images of S. pyogenes , M. catarrhalis and P. acnes biofilms obtained by CLSM. Panels A, C and E show control biofilms, while panels B, D and F show biofilms co-cultured in presence of the probiotic strains by means of transwell inserts. Green = live cells; red = dead cells; 40× magnification
Figure Legend Snippet: Representative images of S. pyogenes , M. catarrhalis and P. acnes biofilms obtained by CLSM. Panels A, C and E show control biofilms, while panels B, D and F show biofilms co-cultured in presence of the probiotic strains by means of transwell inserts. Green = live cells; red = dead cells; 40× magnification

Techniques Used: Confocal Laser Scanning Microscopy, Cell Culture

Inhibition of biofilm formation by cell-free extracts. Data are expressed as mean percentage in respect to biofilm growth control in fresh BHI broth. Black bars = control; grey tone bars = untreated cell-free extract (NT); slanted lines bars = pH neutralized cell-free extract (pH); dotted bars = heat inactivated cell-free extract (TH). * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001
Figure Legend Snippet: Inhibition of biofilm formation by cell-free extracts. Data are expressed as mean percentage in respect to biofilm growth control in fresh BHI broth. Black bars = control; grey tone bars = untreated cell-free extract (NT); slanted lines bars = pH neutralized cell-free extract (pH); dotted bars = heat inactivated cell-free extract (TH). * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Techniques Used: Inhibition

Interaction between S. salivarius 24SMB and S. oralis 89a. Data are expressed as mean absorbance ± standard deviation ( n = 3). Sal = S. salivarius; Sor = S. oralis; ** P ≤ 0.01; *** P ≤ 0.001. Panel A shows the effect of S. oralis 89a on S. salivarius 24SMB biofilm in indirect (transwell) and direct (mixed) contact. Panel B shows the effect of S. salivarius 24SMB on S. oralis 89a biofilm in indirect (transwell) and direct (mixed) contact
Figure Legend Snippet: Interaction between S. salivarius 24SMB and S. oralis 89a. Data are expressed as mean absorbance ± standard deviation ( n = 3). Sal = S. salivarius; Sor = S. oralis; ** P ≤ 0.01; *** P ≤ 0.001. Panel A shows the effect of S. oralis 89a on S. salivarius 24SMB biofilm in indirect (transwell) and direct (mixed) contact. Panel B shows the effect of S. salivarius 24SMB on S. oralis 89a biofilm in indirect (transwell) and direct (mixed) contact

Techniques Used: Standard Deviation

Inhibition of biofilm formation during time. Data are expressed as mean absorbance ± standard deviation (n = 3). Black bars = controls; grey bars = mixed co-cultures; white bars = transwell co-cultures; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001
Figure Legend Snippet: Inhibition of biofilm formation during time. Data are expressed as mean absorbance ± standard deviation (n = 3). Black bars = controls; grey bars = mixed co-cultures; white bars = transwell co-cultures; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Techniques Used: Inhibition, Standard Deviation

Inhibition of pre-formed biofilm during time. Data are expressed as mean percentage in respect to the pre-treatment level ± standard deviation (n = 3). Black bars = pre-treatment level; grey bars = 24 h; white bars = 48 h; dashed bars = 72 h; ctrl = untreated; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001
Figure Legend Snippet: Inhibition of pre-formed biofilm during time. Data are expressed as mean percentage in respect to the pre-treatment level ± standard deviation (n = 3). Black bars = pre-treatment level; grey bars = 24 h; white bars = 48 h; dashed bars = 72 h; ctrl = untreated; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Techniques Used: Inhibition, Standard Deviation

23) Product Images from "Impact of Silver-Containing Wound Dressings on Bacterial Biofilm Viability and Susceptibility to Antibiotics during Prolonged Treatment ▿"

Article Title: Impact of Silver-Containing Wound Dressings on Bacterial Biofilm Viability and Susceptibility to Antibiotics during Prolonged Treatment ▿

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00825-10

SEM images of MRSA biofilms developed on the peg surface before exposure (left) to NCPE, MSN, SCMC, MSAL, and MSPU and after 1 day (middle) and 7 days (right) of the treatment in a daily transfer assay. Bar, 5 μm.
Figure Legend Snippet: SEM images of MRSA biofilms developed on the peg surface before exposure (left) to NCPE, MSN, SCMC, MSAL, and MSPU and after 1 day (middle) and 7 days (right) of the treatment in a daily transfer assay. Bar, 5 μm.

Techniques Used:

Time-dependent effects of silver-containing dressings on biofilm bacterial viability and colony morphology.
Figure Legend Snippet: Time-dependent effects of silver-containing dressings on biofilm bacterial viability and colony morphology.

Techniques Used:

SEM images of P. aeruginosa biofilms developed on the peg surface before exposure (left) to Acticoat nanocrystalline silver on polyethylene mesh (NCPE), Silverlon metallic silver on nylon core (MSN), Aquacel Ag silver carboxymethylcellulose (SCMC), SilverCel
Figure Legend Snippet: SEM images of P. aeruginosa biofilms developed on the peg surface before exposure (left) to Acticoat nanocrystalline silver on polyethylene mesh (NCPE), Silverlon metallic silver on nylon core (MSN), Aquacel Ag silver carboxymethylcellulose (SCMC), SilverCel

Techniques Used:

SEM images of E. coli biofilms developed on the peg surface before exposure (left) to NCPE, MSN, SCMC, MSAL, and MSPU and after 1 day (middle) and 7 days (right) of the treatment in a daily transfer assay. Bar, 5 μm.
Figure Legend Snippet: SEM images of E. coli biofilms developed on the peg surface before exposure (left) to NCPE, MSN, SCMC, MSAL, and MSPU and after 1 day (middle) and 7 days (right) of the treatment in a daily transfer assay. Bar, 5 μm.

Techniques Used:

Antibiotic susceptibilities of biofilms pretreated with silver dressings.
Figure Legend Snippet: Antibiotic susceptibilities of biofilms pretreated with silver dressings.

Techniques Used:

Accumulation of silver in the biofilms.
Figure Legend Snippet: Accumulation of silver in the biofilms.

Techniques Used:

24) Product Images from "The Chromosomal Toxin Gene yafQ Is a Determinant of Multidrug Tolerance for Escherichia coli Growing in a Biofilm ▿"

Article Title: The Chromosomal Toxin Gene yafQ Is a Determinant of Multidrug Tolerance for Escherichia coli Growing in a Biofilm ▿

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00043-09

Stationary-phase planktonic cell populations of Δ yafQ and isogenic parental strain E. coli K-12 BW25113 had similar numbers of cells surviving exposure to cefazolin and tobramycin. Mean log killing was calculated after the biofilms had been exposed
Figure Legend Snippet: Stationary-phase planktonic cell populations of Δ yafQ and isogenic parental strain E. coli K-12 BW25113 had similar numbers of cells surviving exposure to cefazolin and tobramycin. Mean log killing was calculated after the biofilms had been exposed

Techniques Used:

Overexpression of yafQ from a high-copy-number plasmid had no effect on the number of cells in E. coli K-12 BW25113 biofilm populations surviving exposure to doxycycline or rifampin. Mean log killing was calculated from the viable cell counts after the
Figure Legend Snippet: Overexpression of yafQ from a high-copy-number plasmid had no effect on the number of cells in E. coli K-12 BW25113 biofilm populations surviving exposure to doxycycline or rifampin. Mean log killing was calculated from the viable cell counts after the

Techniques Used: Over Expression, Plasmid Preparation

Biofilm populations of the Δ yafQ strain have decreased numbers of cells surviving exposure to cefazolin and tobramycin compared to the numbers of parental E. coli K-12 BW25113 cells. Mean log killing was calculated after the biofilms had been
Figure Legend Snippet: Biofilm populations of the Δ yafQ strain have decreased numbers of cells surviving exposure to cefazolin and tobramycin compared to the numbers of parental E. coli K-12 BW25113 cells. Mean log killing was calculated after the biofilms had been

Techniques Used:

Biofilm formation by wild-type E. coli K-12 BW25113 is similar to that of its isogenic Δ yafQ mutant. (a) Biofilm mean viable cell counts and standard deviations were comparable for the two strains, and this assessment was based on the indicated
Figure Legend Snippet: Biofilm formation by wild-type E. coli K-12 BW25113 is similar to that of its isogenic Δ yafQ mutant. (a) Biofilm mean viable cell counts and standard deviations were comparable for the two strains, and this assessment was based on the indicated

Techniques Used: Mutagenesis

Biofilm populations of the Δ yafQ strain and isogenic parental strain E. coli K-12 BW25113 had similar numbers of cells surviving exposure to doxycycline and rifampin. Mean log killing was calculated from the viable cell counts after the biofilms
Figure Legend Snippet: Biofilm populations of the Δ yafQ strain and isogenic parental strain E. coli K-12 BW25113 had similar numbers of cells surviving exposure to doxycycline and rifampin. Mean log killing was calculated from the viable cell counts after the biofilms

Techniques Used:

Overexpression of yafQ from a high-copy-number plasmid increased the number of cells in E. coli K-12 BW25113 biofilm populations surviving exposure to bactericidal concentrations of cefazolin and tobramycin. Mean log killing was calculated from the viable
Figure Legend Snippet: Overexpression of yafQ from a high-copy-number plasmid increased the number of cells in E. coli K-12 BW25113 biofilm populations surviving exposure to bactericidal concentrations of cefazolin and tobramycin. Mean log killing was calculated from the viable

Techniques Used: Over Expression, Plasmid Preparation

25) Product Images from "The use of microscopy and three-dimensional visualization to evaluate the structure of microbial biofilms cultivated in the Calgary Biofilm Device"

Article Title: The use of microscopy and three-dimensional visualization to evaluate the structure of microbial biofilms cultivated in the Calgary Biofilm Device

Journal: Biological Procedures Online

doi: 10.1251/bpo127

3D visualization of microbial biofilms using amira®. A. A 2D average of an image z-stack for a P. aeruginosa PA14 biofilm stained with AO and TRITC-ConA. B and C. Isosurface rendering of the 3D volume data sets for AO and TRITC-ConA, respectively, extrapolated from the image z-stacks used to create the image in panel A. D. The merged, isosurface rendered 3D volume data set for AO and TRITC-ConA. E to G. Volume rendering corresponding to the data sets presented in panels B to D. H. A 2D average of an image z-stack for a C. tropicalis 99916 biofilm stained with the Live/Dead® cell viability kit. I and J. Isosurface rendering of the 3D volume data sets for Syto-9 and PI respectively, extrapolated from the image z-stacks used to create the image in panel H. K. The merged, isosurface rendered 3D volume data set for Syto-9 and PI. L to M. Volume rendering corresponding to the data sets presented in panels I to K. O and P. The oblique view of the P. aeruginosa PA14 biofilm pictured in panel A visualized using isosurface and volume rendering, respectively. Q and R. The oblique view of the C. tropicalis 99916 biofilm pictured in panel H visualized using isosurface and volume rendering, respectively. Each 2D image panel or 3D model represents a square surface area of approximately 238 × 238 μm.
Figure Legend Snippet: 3D visualization of microbial biofilms using amira®. A. A 2D average of an image z-stack for a P. aeruginosa PA14 biofilm stained with AO and TRITC-ConA. B and C. Isosurface rendering of the 3D volume data sets for AO and TRITC-ConA, respectively, extrapolated from the image z-stacks used to create the image in panel A. D. The merged, isosurface rendered 3D volume data set for AO and TRITC-ConA. E to G. Volume rendering corresponding to the data sets presented in panels B to D. H. A 2D average of an image z-stack for a C. tropicalis 99916 biofilm stained with the Live/Dead® cell viability kit. I and J. Isosurface rendering of the 3D volume data sets for Syto-9 and PI respectively, extrapolated from the image z-stacks used to create the image in panel H. K. The merged, isosurface rendered 3D volume data set for Syto-9 and PI. L to M. Volume rendering corresponding to the data sets presented in panels I to K. O and P. The oblique view of the P. aeruginosa PA14 biofilm pictured in panel A visualized using isosurface and volume rendering, respectively. Q and R. The oblique view of the C. tropicalis 99916 biofilm pictured in panel H visualized using isosurface and volume rendering, respectively. Each 2D image panel or 3D model represents a square surface area of approximately 238 × 238 μm.

Techniques Used: Staining

An overview for using the CBD for the purpose of microscopy and 3D visualization of microbial biofilms. A. To begin, fresh subcultures of the microbial strain were grown on the appropriate agar medium. B. Using a cotton swab, colonies from a fresh second subculture were suspended in broth medium to match a 1.0 McFarland standard. This was diluted 30-fold in broth to create the inoculum for the CBD. C. The peg lid of the CBD was either inserted into a microtiter plate (containing 150 μL of inoculum in each well) or a corrugated trough (with 22 mL of inoculum inside). The inoculated devices were placed in a humidified incubator on a gyrorotary shaker or platform rocker, respectively. D. After cultivation, biofilms were rinsed with saline to remove loosely adherent cells. E. Pegs were removed from the CBD using pliers and the biofilms then were enumerated by viable cell counting. A second set of pegs was removed for examination by microscopy. There is an option to expose biofilms to an array of antimicrobial agents or other test conditions and then to remove a second set of pegs for microscopy. F. For SEM, pegs were first fixed and then dehydrated, which was carried out using 1 of 2 protocols. G. The fixed samples were mounted on stubs using epoxy resin, dried, and then sputter coated with gold-palladium. H. The biofilms were then examined by SEM, and the resulting images were contrast enhanced. I. For CLSM, pegs were immersed in the appropriate stain and then placed in 2 drops of 0.9% saline on a glass coverslip. J. Images of the biofilms were captured using CLSM, and the instrument software was used to generate 2D averages of image z-stacks. K. The z-stacks were imported into amira™ for advanced image processing and 3D visualization.
Figure Legend Snippet: An overview for using the CBD for the purpose of microscopy and 3D visualization of microbial biofilms. A. To begin, fresh subcultures of the microbial strain were grown on the appropriate agar medium. B. Using a cotton swab, colonies from a fresh second subculture were suspended in broth medium to match a 1.0 McFarland standard. This was diluted 30-fold in broth to create the inoculum for the CBD. C. The peg lid of the CBD was either inserted into a microtiter plate (containing 150 μL of inoculum in each well) or a corrugated trough (with 22 mL of inoculum inside). The inoculated devices were placed in a humidified incubator on a gyrorotary shaker or platform rocker, respectively. D. After cultivation, biofilms were rinsed with saline to remove loosely adherent cells. E. Pegs were removed from the CBD using pliers and the biofilms then were enumerated by viable cell counting. A second set of pegs was removed for examination by microscopy. There is an option to expose biofilms to an array of antimicrobial agents or other test conditions and then to remove a second set of pegs for microscopy. F. For SEM, pegs were first fixed and then dehydrated, which was carried out using 1 of 2 protocols. G. The fixed samples were mounted on stubs using epoxy resin, dried, and then sputter coated with gold-palladium. H. The biofilms were then examined by SEM, and the resulting images were contrast enhanced. I. For CLSM, pegs were immersed in the appropriate stain and then placed in 2 drops of 0.9% saline on a glass coverslip. J. Images of the biofilms were captured using CLSM, and the instrument software was used to generate 2D averages of image z-stacks. K. The z-stacks were imported into amira™ for advanced image processing and 3D visualization.

Techniques Used: Microscopy, Cell Counting, Confocal Laser Scanning Microscopy, Staining, Software

SEM of bacterial biofilms grown in the CBD. For the sake of comparison, the biofilms were grown in either rich or minimal medium (as summarized in Table 1 ) and then fixed using 1 of 2 different protocols. These micrographs were chosen to illustrate that medium composition has an impact on the capacity of bacteria to form biofilms, which further varies between genus, species and strains. Moreover, the choice of fixing protocols influences how well microstructures may be preserved, which may impact on the interpretation of SEM data.
Figure Legend Snippet: SEM of bacterial biofilms grown in the CBD. For the sake of comparison, the biofilms were grown in either rich or minimal medium (as summarized in Table 1 ) and then fixed using 1 of 2 different protocols. These micrographs were chosen to illustrate that medium composition has an impact on the capacity of bacteria to form biofilms, which further varies between genus, species and strains. Moreover, the choice of fixing protocols influences how well microstructures may be preserved, which may impact on the interpretation of SEM data.

Techniques Used:

CLSM of AO stained bacterial biofilms grown in the CBD. The images on the left are 2D averages of image z-stacks, whereas the images on the right are isosurface rendered 3D visualizations of the same data set (prepared using Leica® Confocal Software). These data sets illustrate that mature biofilms may adopt a number of structures that are distinct from the archetypal “stalk and mushroom” microcolony structures that are well characterized for P. aeruginosa. Each panel represents a square surface area of approximately 238 × 238 μm.
Figure Legend Snippet: CLSM of AO stained bacterial biofilms grown in the CBD. The images on the left are 2D averages of image z-stacks, whereas the images on the right are isosurface rendered 3D visualizations of the same data set (prepared using Leica® Confocal Software). These data sets illustrate that mature biofilms may adopt a number of structures that are distinct from the archetypal “stalk and mushroom” microcolony structures that are well characterized for P. aeruginosa. Each panel represents a square surface area of approximately 238 × 238 μm.

Techniques Used: Confocal Laser Scanning Microscopy, Staining, Software

CLSM of Live/Dead® (Syto-9 and PI) stained microbial biofilms grown in the CBD. Cell death is a normal part of biofilm development (the extent of which may vary) and therefore dead biomass constitutes a portion of every biofilm. Each panel represents a square surface area of approximately 238 × 238 μm.
Figure Legend Snippet: CLSM of Live/Dead® (Syto-9 and PI) stained microbial biofilms grown in the CBD. Cell death is a normal part of biofilm development (the extent of which may vary) and therefore dead biomass constitutes a portion of every biofilm. Each panel represents a square surface area of approximately 238 × 238 μm.

Techniques Used: Confocal Laser Scanning Microscopy, Staining

TRITC-conjugated lectin staining of C. tropicalis and B. cenocepacia biofilms. A. An overlay image of a C. tropicalis biofilm that was stained with AO (green emission) and TRITC-ConA (red emission). B. AO staining of the section illustrated in panel A. C. TRITC-ConA staining of the section illustrated in panel A. D. An overlay image of a B. cenocepacia biofilm that was stained with Syto-9 and TRITC-PNA. E. Syto-9 staining of the section illustrated in panel D. F. TRITC-PNA staining of the section illustrated in panel D. Each panel represents a square surface area of approximately 238 × 238 μm.
Figure Legend Snippet: TRITC-conjugated lectin staining of C. tropicalis and B. cenocepacia biofilms. A. An overlay image of a C. tropicalis biofilm that was stained with AO (green emission) and TRITC-ConA (red emission). B. AO staining of the section illustrated in panel A. C. TRITC-ConA staining of the section illustrated in panel A. D. An overlay image of a B. cenocepacia biofilm that was stained with Syto-9 and TRITC-PNA. E. Syto-9 staining of the section illustrated in panel D. F. TRITC-PNA staining of the section illustrated in panel D. Each panel represents a square surface area of approximately 238 × 238 μm.

Techniques Used: Staining

26) Product Images from "Staphylococcus epidermidis in Orthopedic Device Infections: The Role of Bacterial Internalization in Human Osteoblasts and Biofilm Formation"

Article Title: Staphylococcus epidermidis in Orthopedic Device Infections: The Role of Bacterial Internalization in Human Osteoblasts and Biofilm Formation

Journal: PLoS ONE

doi: 10.1371/journal.pone.0067240

Evaluation of biofilm-formation ability of S. epidermidis clinical isolates. A. Kinetics of early biofilm formation was assayed by the Biofilm Ring Test method for the reference strain NCTC11047 and for infective (n = 15) and colonizing (n = 22) S. epidermidis strains. B. Quantification of mature biofilm formation after 24 and 48 h by the crystal violet staining method for the S. epidermidis reference strain NCTC11047 and 37 clinical isolates.
Figure Legend Snippet: Evaluation of biofilm-formation ability of S. epidermidis clinical isolates. A. Kinetics of early biofilm formation was assayed by the Biofilm Ring Test method for the reference strain NCTC11047 and for infective (n = 15) and colonizing (n = 22) S. epidermidis strains. B. Quantification of mature biofilm formation after 24 and 48 h by the crystal violet staining method for the S. epidermidis reference strain NCTC11047 and 37 clinical isolates.

Techniques Used: Staining

27) Product Images from "Mycobacterium avium Biofilm Attenuates Mononuclear Phagocyte Function by Triggering Hyperstimulation and Apoptosis during Early Infection"

Article Title: Mycobacterium avium Biofilm Attenuates Mononuclear Phagocyte Function by Triggering Hyperstimulation and Apoptosis during Early Infection

Journal: Infection and Immunity

doi: 10.1128/IAI.00820-13

THP-1 cells rapidly undergo apoptosis after exposure to M. avium subsp. hominissuis A5 biofilm. (A to F) Microscopy showing THP-1 cells at 5 min (A), 15 min (B), 25 min (C and D), and 40 min (E and F). (G) THP-1 cells were placed on top of established M. avium subsp. hominissuis A5 biofilm, and the TUNEL assay was performed at 0, 0.5, 24, and 48 h. In each field of view, apoptotic cells were counted microscopically and divided by the total number of counted cells to attain percentage of apoptosis. Fifty fields of view were counted for each time point and averaged. The bars represent the mean of each time point ± the standard deviation.
Figure Legend Snippet: THP-1 cells rapidly undergo apoptosis after exposure to M. avium subsp. hominissuis A5 biofilm. (A to F) Microscopy showing THP-1 cells at 5 min (A), 15 min (B), 25 min (C and D), and 40 min (E and F). (G) THP-1 cells were placed on top of established M. avium subsp. hominissuis A5 biofilm, and the TUNEL assay was performed at 0, 0.5, 24, and 48 h. In each field of view, apoptotic cells were counted microscopically and divided by the total number of counted cells to attain percentage of apoptosis. Fifty fields of view were counted for each time point and averaged. The bars represent the mean of each time point ± the standard deviation.

Techniques Used: Microscopy, TUNEL Assay, Standard Deviation

THP-1 cells are stimulated during exposure to M. avium subsp. hominissuis biofilms. (A) THP-1 cells were added on top of M. avium subsp. hominissuis strain A5 and 104 14-day-old biofilms (equal inoculum levels), and supernatant was collected at 0.5, 1, 2, 4, 24, and 48 h. TNF-α ELISA was carried out for each time point for each strain. (B) Filter-sterilized supernatants from 14-day-old biofilms were added at a 10% (vol/vol) concentration in RPMI 1640 plus 10% FBS with THP-1 cells for 4 h. Culture supernatant was then collected, and TNF-α ELISA was performed. Abbreviations: NS, not stimulated; 104 S, M. avium subsp. hominissuis 104 supernatant; A5 S, M. avium subsp. hominissuis A5 supernatant. (C) UV-killed M. avium subsp. hominissuis A5 biofilms (A5 BF UV) were compared side by side with non-UV-treated biofilms (A5 BF) and planktonic bacteria that were immobilized on the bottom surface of the plate via centrifugation (A5 plank) for TNF-α production by THP-1 cells 4 h after being placed on top of the bacteria. (D) THP-1 cells were placed on top of M. avium subsp. hominissuis A5 biofilm, and O 2 − was assessed spectrophotometrically as described in Materials and Methods. (E) THP-1 cells were placed on top of M. avium subsp. hominissuis A5 biofilm, and nitric oxide (NO) was assessed using the Griess reagent system. Abbreviations for panels D and E: neg, negative control for respective assay; THP-1, THP-1 cells placed in empty wells with no bacteria present; THP-1 A5, THP-1 cells placed in wells containing M. avium subsp. hominissuis A5 biofilm. Bars represent means ± standard deviations. *, P
Figure Legend Snippet: THP-1 cells are stimulated during exposure to M. avium subsp. hominissuis biofilms. (A) THP-1 cells were added on top of M. avium subsp. hominissuis strain A5 and 104 14-day-old biofilms (equal inoculum levels), and supernatant was collected at 0.5, 1, 2, 4, 24, and 48 h. TNF-α ELISA was carried out for each time point for each strain. (B) Filter-sterilized supernatants from 14-day-old biofilms were added at a 10% (vol/vol) concentration in RPMI 1640 plus 10% FBS with THP-1 cells for 4 h. Culture supernatant was then collected, and TNF-α ELISA was performed. Abbreviations: NS, not stimulated; 104 S, M. avium subsp. hominissuis 104 supernatant; A5 S, M. avium subsp. hominissuis A5 supernatant. (C) UV-killed M. avium subsp. hominissuis A5 biofilms (A5 BF UV) were compared side by side with non-UV-treated biofilms (A5 BF) and planktonic bacteria that were immobilized on the bottom surface of the plate via centrifugation (A5 plank) for TNF-α production by THP-1 cells 4 h after being placed on top of the bacteria. (D) THP-1 cells were placed on top of M. avium subsp. hominissuis A5 biofilm, and O 2 − was assessed spectrophotometrically as described in Materials and Methods. (E) THP-1 cells were placed on top of M. avium subsp. hominissuis A5 biofilm, and nitric oxide (NO) was assessed using the Griess reagent system. Abbreviations for panels D and E: neg, negative control for respective assay; THP-1, THP-1 cells placed in empty wells with no bacteria present; THP-1 A5, THP-1 cells placed in wells containing M. avium subsp. hominissuis A5 biofilm. Bars represent means ± standard deviations. *, P

Techniques Used: Enzyme-linked Immunosorbent Assay, Concentration Assay, Centrifugation, Negative Control

Blocking TNF-α reduces apoptosis of THP-1 cells during M. avium subsp. hominissuis A5 biofilm exposure. THP-1 cells were placed on top of established M. avium subsp. hominissuis A5 biofilms with or without anti-TNF-R1 (1:10 dilution of a hybridoma suspension) or anti-TNF-α (10 μg/ml). (A) TUNEL assay was performed at 2 and 6 h after exposure. The ratio of apoptotic to total cells was counted in 50 fields of view microscopically and was averaged for each time point. (B) THP-1 cells were lysed at 24 h after exposure, and wells were resuspended, diluted, and plated for CFU of M. avium subsp. hominissuis . Bars represent the mean ± standard deviation. *, P
Figure Legend Snippet: Blocking TNF-α reduces apoptosis of THP-1 cells during M. avium subsp. hominissuis A5 biofilm exposure. THP-1 cells were placed on top of established M. avium subsp. hominissuis A5 biofilms with or without anti-TNF-R1 (1:10 dilution of a hybridoma suspension) or anti-TNF-α (10 μg/ml). (A) TUNEL assay was performed at 2 and 6 h after exposure. The ratio of apoptotic to total cells was counted in 50 fields of view microscopically and was averaged for each time point. (B) THP-1 cells were lysed at 24 h after exposure, and wells were resuspended, diluted, and plated for CFU of M. avium subsp. hominissuis . Bars represent the mean ± standard deviation. *, P

Techniques Used: Blocking Assay, TUNEL Assay, Standard Deviation

The number of CFU of M. avium subsp. hominissuis A5 biofilm is unaffected by the addition of THP-1 cells, even when preactivated or cocultured with NK cells. (A) THP-1 cells were added on top of established M. avium subsp. hominissuis A5 biofilms for 24 and 72 h. In some instances, THP-1 cells were first prestimulated with 50 ng/ml of TNF-α or 100 ng/ml of IFN-γ. At each time point, wells were resuspended, diluted, and plated to obtain CFU/well of M. avium subsp. hominissuis A5. (B) Suspensions of THP-1 cells alone, NK cells alone, or a coculture of the two cell types were added on top of established M. avium subsp. hominissuis A5 biofilms for 24 and 48 h before CFU enumeration. (C) Coculture of NK cells and THP-1 cells infected with planktonic M. avium subsp. hominissuis demonstrating NK cell-assisted bacterial killing after 4 days. (D) Infection of THP-1 with planktonic M. avium subsp. hominissuis A5 and two M. avium subsp. hominissuis A5-derived biofilm-deficient mutants (6H9 and 5G4), demonstrating that planktonic M. avium subsp. hominissuis can grow inside THP-1 cells. For both panels A and B, all comparisons to the M. avium subsp. hominissuis A5 control (no host cells added) for each time point were not significant. Bars represent means ± standard deviations. **, P
Figure Legend Snippet: The number of CFU of M. avium subsp. hominissuis A5 biofilm is unaffected by the addition of THP-1 cells, even when preactivated or cocultured with NK cells. (A) THP-1 cells were added on top of established M. avium subsp. hominissuis A5 biofilms for 24 and 72 h. In some instances, THP-1 cells were first prestimulated with 50 ng/ml of TNF-α or 100 ng/ml of IFN-γ. At each time point, wells were resuspended, diluted, and plated to obtain CFU/well of M. avium subsp. hominissuis A5. (B) Suspensions of THP-1 cells alone, NK cells alone, or a coculture of the two cell types were added on top of established M. avium subsp. hominissuis A5 biofilms for 24 and 48 h before CFU enumeration. (C) Coculture of NK cells and THP-1 cells infected with planktonic M. avium subsp. hominissuis demonstrating NK cell-assisted bacterial killing after 4 days. (D) Infection of THP-1 with planktonic M. avium subsp. hominissuis A5 and two M. avium subsp. hominissuis A5-derived biofilm-deficient mutants (6H9 and 5G4), demonstrating that planktonic M. avium subsp. hominissuis can grow inside THP-1 cells. For both panels A and B, all comparisons to the M. avium subsp. hominissuis A5 control (no host cells added) for each time point were not significant. Bars represent means ± standard deviations. **, P

Techniques Used: Infection, Derivative Assay

28) Product Images from "Hyperadherence of Pseudomonas taiwanensis VLB120ΔC increases productivity of (S)‐styrene oxide formation"

Article Title: Hyperadherence of Pseudomonas taiwanensis VLB120ΔC increases productivity of (S)‐styrene oxide formation

Journal: Microbial Biotechnology

doi: 10.1111/1751-7915.12378

Biofilm dry weight ( BDW ) and viable cells in biofilms determined via the resazurin and eGFP assay. Values are given as a ratio of the quantities of the mutant Pseudomonas taiwanensis VLB 120ΔCe GFP Δ04710 to the parent strain P. taiwanensis VLB 120ΔCe GFP . A. Biofilms after 48 h of standard cultivation under segmented flow conditions. Errors bars represent standard deviation of triplicates. B. Biofilms after 336 h of biotransformation under segmented flow conditions. Error bars represent standard deviation of five replicates.
Figure Legend Snippet: Biofilm dry weight ( BDW ) and viable cells in biofilms determined via the resazurin and eGFP assay. Values are given as a ratio of the quantities of the mutant Pseudomonas taiwanensis VLB 120ΔCe GFP Δ04710 to the parent strain P. taiwanensis VLB 120ΔCe GFP . A. Biofilms after 48 h of standard cultivation under segmented flow conditions. Errors bars represent standard deviation of triplicates. B. Biofilms after 336 h of biotransformation under segmented flow conditions. Error bars represent standard deviation of five replicates.

Techniques Used: Mutagenesis, Flow Cytometry, Standard Deviation

Attached and detached biofilm biomass of Pseudomonas taiwanensis VLB 120ΔCe GFP Δ04710 and the control strain P. taiwanensis VLB 120ΔCe GFP grown in silicone tubes applying single‐phase aqueous flow rates of 100 μl min −1 and 500 μl min −1 or two‐phase aqueous‐air segmented flow of 100 μl min −1 medium and 100 μl min −1 air. Errors bars represent standard deviation of triplicates. A. Total biofilm dry weight ( BDW ) of biofilms grown in 40 cm long silicone tubings. B. Detached biofilm biomass during continuous biofilm cultivation. One data point reflects the amount of biomass in 10 ml effluent.
Figure Legend Snippet: Attached and detached biofilm biomass of Pseudomonas taiwanensis VLB 120ΔCe GFP Δ04710 and the control strain P. taiwanensis VLB 120ΔCe GFP grown in silicone tubes applying single‐phase aqueous flow rates of 100 μl min −1 and 500 μl min −1 or two‐phase aqueous‐air segmented flow of 100 μl min −1 medium and 100 μl min −1 air. Errors bars represent standard deviation of triplicates. A. Total biofilm dry weight ( BDW ) of biofilms grown in 40 cm long silicone tubings. B. Detached biofilm biomass during continuous biofilm cultivation. One data point reflects the amount of biomass in 10 ml effluent.

Techniques Used: Flow Cytometry, Standard Deviation

29) Product Images from "Modulation of the Substitution Pattern of 5-Aryl-2-Aminoimidazoles Allows Fine-Tuning of Their Antibiofilm Activity Spectrum and Toxicity"

Article Title: Modulation of the Substitution Pattern of 5-Aryl-2-Aminoimidazoles Allows Fine-Tuning of Their Antibiofilm Activity Spectrum and Toxicity

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00035-16

Structures of 5-Ar-2-AI-based compounds whose activities against a broad panel of monospecies and mixed-species biofilm models were tested in this study.
Figure Legend Snippet: Structures of 5-Ar-2-AI-based compounds whose activities against a broad panel of monospecies and mixed-species biofilm models were tested in this study.

Techniques Used:

Synthesis and structures of eight novel N 1-,2 N -disubstituted 5-Ar-2-AIs tested against monospecies and mixed-species biofilms. MeOH, methanol; rt, room temperature. Percentages indicate compound yield.
Figure Legend Snippet: Synthesis and structures of eight novel N 1-,2 N -disubstituted 5-Ar-2-AIs tested against monospecies and mixed-species biofilms. MeOH, methanol; rt, room temperature. Percentages indicate compound yield.

Techniques Used:

30) Product Images from "The Behavior of Staphylococcus aureus Dual-Species Biofilms Treated with Bacteriophage phiIPLA-RODI Depends on the Accompanying Microorganism"

Article Title: The Behavior of Staphylococcus aureus Dual-Species Biofilms Treated with Bacteriophage phiIPLA-RODI Depends on the Accompanying Microorganism

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.02821-16

Effect of phage phiIPLA-RODI on 5 h (A) and 24 h (B) biofilms formed by S. aureus IPLA16- L. plantarum 55-1 and S. aureus IPLA16- E. faecium MMRA after 4 h of treatment with 10 9 PFU/well in SM buffer. Values represent the means ± standard deviations
Figure Legend Snippet: Effect of phage phiIPLA-RODI on 5 h (A) and 24 h (B) biofilms formed by S. aureus IPLA16- L. plantarum 55-1 and S. aureus IPLA16- E. faecium MMRA after 4 h of treatment with 10 9 PFU/well in SM buffer. Values represent the means ± standard deviations

Techniques Used:

Scanning electron micrographs of S. aureus dual-species biofilms. Images correspond to 5-h biofilms formed by S. aureus IPLA16 with E. faecium MMRA (A and B), L. plantarum 55-1 (C and D), L. pentosus A1 (E and F), or L. pentosus B1 (G and H) following
Figure Legend Snippet: Scanning electron micrographs of S. aureus dual-species biofilms. Images correspond to 5-h biofilms formed by S. aureus IPLA16 with E. faecium MMRA (A and B), L. plantarum 55-1 (C and D), L. pentosus A1 (E and F), or L. pentosus B1 (G and H) following

Techniques Used:

Propagation of phage phiIPLA-RODI on 5-h dual-species biofilms of S. aureus IPLA16 with other bacteria treated for 18 h with 10 9 PFU/well or 10 6 PFU/well. The accompanying species tested were E. faecium MMRA (A), L. plantarum 55-1 (B), L. pentosus A1
Figure Legend Snippet: Propagation of phage phiIPLA-RODI on 5-h dual-species biofilms of S. aureus IPLA16 with other bacteria treated for 18 h with 10 9 PFU/well or 10 6 PFU/well. The accompanying species tested were E. faecium MMRA (A), L. plantarum 55-1 (B), L. pentosus A1

Techniques Used:

CLSM images of S. aureus IPLA16 and E. faecium MMRA (A and B), L. plantarum 55-1 (C and D), L. pentosus A1 (E and F), or L. pentosus B1 (G and H) dual-species biofilms. Images correspond to 5-h biofilms untreated (A, C, E, and G) or treated with phiIPLA-RODI
Figure Legend Snippet: CLSM images of S. aureus IPLA16 and E. faecium MMRA (A and B), L. plantarum 55-1 (C and D), L. pentosus A1 (E and F), or L. pentosus B1 (G and H) dual-species biofilms. Images correspond to 5-h biofilms untreated (A, C, E, and G) or treated with phiIPLA-RODI

Techniques Used: Confocal Laser Scanning Microscopy

Biofilms formed by S. aureus IPLA16 with L. plantarum 55-1, E. faecium MMRA, L. pentosus A1, and L. pentosus B1 after incubating 5 h at 32°C or 37°C. (A) Biomass of monospecies biofilms formed by S. aureus IPLA16 (white bars), accompanying
Figure Legend Snippet: Biofilms formed by S. aureus IPLA16 with L. plantarum 55-1, E. faecium MMRA, L. pentosus A1, and L. pentosus B1 after incubating 5 h at 32°C or 37°C. (A) Biomass of monospecies biofilms formed by S. aureus IPLA16 (white bars), accompanying

Techniques Used:

Bacterial counts of 5-h biofilms formed by S. aureus IPLA16 with other species under proliferation conditions and treated with phage phiIPLA-RODI. Values represent the means ± standard deviations of two independent biological replicates. The accompanying
Figure Legend Snippet: Bacterial counts of 5-h biofilms formed by S. aureus IPLA16 with other species under proliferation conditions and treated with phage phiIPLA-RODI. Values represent the means ± standard deviations of two independent biological replicates. The accompanying

Techniques Used:

31) Product Images from "Metal Ions May Suppress or Enhance Cellular Differentiation in Candida albicans and Candida tropicalis Biofilms ▿ Biofilms ▿ †"

Article Title: Metal Ions May Suppress or Enhance Cellular Differentiation in Candida albicans and Candida tropicalis Biofilms ▿ Biofilms ▿ †

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.02711-06

Cultivation of C. tropicalis 99916 biofilms in SeO 3 2− and CrO 4 2− decreases the resistance of the fungal community to amphotericin B and Cu 2+ , respectively. In these experiments, C. tropicalis biofilms were grown on pegs in the
Figure Legend Snippet: Cultivation of C. tropicalis 99916 biofilms in SeO 3 2− and CrO 4 2− decreases the resistance of the fungal community to amphotericin B and Cu 2+ , respectively. In these experiments, C. tropicalis biofilms were grown on pegs in the

Techniques Used:

C. albicans 3153A and C. tropicalis 99916 undergo multiple shifts in growth rate and biofilm community structure when cultivated in a concentration gradient of divalent mercury cations (Hg 2+ ). Biofilms were grown in RPMI 1640 plus l -glutamine
Figure Legend Snippet: C. albicans 3153A and C. tropicalis 99916 undergo multiple shifts in growth rate and biofilm community structure when cultivated in a concentration gradient of divalent mercury cations (Hg 2+ ). Biofilms were grown in RPMI 1640 plus l -glutamine

Techniques Used: Concentration Assay

Metal ions may promote or inhibit cellular differentiation during biofilm growth of C. albicans 3153A and C. tropicalis 99916. Biofilms were grown in RPMI 1640 plus l -glutamine plus 0.165 M MOPS in microtiter plates for 48 h at 35°C on a gyratory
Figure Legend Snippet: Metal ions may promote or inhibit cellular differentiation during biofilm growth of C. albicans 3153A and C. tropicalis 99916. Biofilms were grown in RPMI 1640 plus l -glutamine plus 0.165 M MOPS in microtiter plates for 48 h at 35°C on a gyratory

Techniques Used: Cell Differentiation

C. tropicalis 99916 forms biofilm communities with characteristic 3D structure that may be influenced by metal ions. Here, the heavy metal ions Zn 2+ and CrO 4 2− influenced the maturation of C. tropicalis communities at an intermediate stage
Figure Legend Snippet: C. tropicalis 99916 forms biofilm communities with characteristic 3D structure that may be influenced by metal ions. Here, the heavy metal ions Zn 2+ and CrO 4 2− influenced the maturation of C. tropicalis communities at an intermediate stage

Techniques Used:

Many metal ions (Co 2+ , Cu 2+ , Ag + , Cd 2+ , Hg 2+ , Pb 2+ , AsO 2 − , and SeO 3 2− ) inhibited hyphal formation by C. tropicalis 99916 at an intermediate stage of biofilm development. In every case, treating
Figure Legend Snippet: Many metal ions (Co 2+ , Cu 2+ , Ag + , Cd 2+ , Hg 2+ , Pb 2+ , AsO 2 − , and SeO 3 2− ) inhibited hyphal formation by C. tropicalis 99916 at an intermediate stage of biofilm development. In every case, treating

Techniques Used: Allele-specific Oligonucleotide

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Tube Formation Assay:

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Cell Culture:

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Incubation:

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Article Snippet: .. Sample preparation and FC Suspensions of reference species and biofilms grown indoors or sampled in the field were sonicated (45 kHz 60 W, VWR Ultrasonic Cleaner) for 1 min to break up colonies, filtered through 50 μm filters (Partec) and immediately fixed (0.01% paraformaldehyde and 0.1% glutaraldehyde (w/v, stock in tap water)) at 4 °C overnight. .. On the basis of the light absorption and fluorescence spectra of the cultured strains and the sampled biofilms measured with a plate reader, dichroic splitters and filters for the Beckman Coulter Gallios flow cytometer (using 405, 488 and 638 nm lasers) were selected to cover fluorescence emission from 425 to > 755 nm ( ).

Article Title: Synergistic effects of heat and antibiotics on Pseudomonas aeruginosa biofilms
Article Snippet: .. To disrupt the biofilms and re-suspend the bacteria in a homogenous solution for serial dilution, each recovery plate with biofilm-covered pegs was sonicated for 10 min at 45 kHz in a VWR Symphony 9.5 L sonicator (Radnor, PA, USA). .. The sonicated recovery plate suspensions were serially diluted tenfold in a 96-well flat-bottom culture plate.

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    Avantor biofilms
    Biofilms, supplied by Avantor, used in various techniques. Bioz Stars score: 94/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biofilms/product/Avantor
    Average 94 stars, based on 22 article reviews
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    biofilms - by Bioz Stars, 2020-05
    94/100 stars
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