Structured Review

BioTek Instruments biofilm
The activities of levofloxacin and Cath-A against the established biofilms of P. aeruginisa ( a ) and A . baumannii ( b ) at different concentrations (4–256 µg/mL). The antibiofilm effect of the peptide and levofloxacin was assessed by cv staining after 24 h of incubation. <t>Biofilm</t> formation was determined by measuring the absorbance at 595 nm using a microplate reader. Controls were untreated inoculated bacteria in Mueller Hinton (MH) broth. Data are shown as the mean ± SD of three independent tests. Significant different values were (# p
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1) Product Images from "A Recombinant Snake Cathelicidin Derivative Peptide: Antibiofilm Properties and Expression in Escherichia coli"

Article Title: A Recombinant Snake Cathelicidin Derivative Peptide: Antibiofilm Properties and Expression in Escherichia coli

Journal: Biomolecules

doi: 10.3390/biom8040118

The activities of levofloxacin and Cath-A against the established biofilms of P. aeruginisa ( a ) and A . baumannii ( b ) at different concentrations (4–256 µg/mL). The antibiofilm effect of the peptide and levofloxacin was assessed by cv staining after 24 h of incubation. Biofilm formation was determined by measuring the absorbance at 595 nm using a microplate reader. Controls were untreated inoculated bacteria in Mueller Hinton (MH) broth. Data are shown as the mean ± SD of three independent tests. Significant different values were (# p
Figure Legend Snippet: The activities of levofloxacin and Cath-A against the established biofilms of P. aeruginisa ( a ) and A . baumannii ( b ) at different concentrations (4–256 µg/mL). The antibiofilm effect of the peptide and levofloxacin was assessed by cv staining after 24 h of incubation. Biofilm formation was determined by measuring the absorbance at 595 nm using a microplate reader. Controls were untreated inoculated bacteria in Mueller Hinton (MH) broth. Data are shown as the mean ± SD of three independent tests. Significant different values were (# p

Techniques Used: Staining, Incubation

2) Product Images from "Cariogenicity induced by commercial carbonated beverages in an experimental biofilm-caries model"

Article Title: Cariogenicity induced by commercial carbonated beverages in an experimental biofilm-caries model

Journal: European Journal of Dentistry

doi: 10.4103/ejd.ejd_188_17

Effect of the different beverages tested on the outcomes of the study: Demineralization on enamel (A) and dentin (B). Bars represent mean percentage of surface hardness loss of the slabs. (II) Biomass induced by each experimental condition. Enamel (A) and dentin (B). Bars represent mean biomass (mg) obtained after exposure to each experimental condition. (III) Viable Streptococcus mutans cells in the biofilms formed on enamel (A) and on dentin (B). Bacterial cells retrieved from each biofilm exposed to test solutions were seeded on plates, counted and expressed as CFU/biofilm dry weight (mg). (IV) Insoluble extracellular polysaccharides produced by the biofilms in response to carbonated beverages. Enamel (A) and dentin (B) biofilms by the different conditions were measured and expressed as mg/mg of biofilm. Bars indicate mean values for each treatment. Error bars indicate standard deviation ( n = 6). Different letters represent significant differences among treatments ( P
Figure Legend Snippet: Effect of the different beverages tested on the outcomes of the study: Demineralization on enamel (A) and dentin (B). Bars represent mean percentage of surface hardness loss of the slabs. (II) Biomass induced by each experimental condition. Enamel (A) and dentin (B). Bars represent mean biomass (mg) obtained after exposure to each experimental condition. (III) Viable Streptococcus mutans cells in the biofilms formed on enamel (A) and on dentin (B). Bacterial cells retrieved from each biofilm exposed to test solutions were seeded on plates, counted and expressed as CFU/biofilm dry weight (mg). (IV) Insoluble extracellular polysaccharides produced by the biofilms in response to carbonated beverages. Enamel (A) and dentin (B) biofilms by the different conditions were measured and expressed as mg/mg of biofilm. Bars indicate mean values for each treatment. Error bars indicate standard deviation ( n = 6). Different letters represent significant differences among treatments ( P

Techniques Used: Produced, Standard Deviation

Acidogenicity from Streptococcus mutans biofilms formed on enamel (A) and dentin (B), exposed to tested carbonated beverages. Plot shows pH of the culture medium. Each point in the plot represents mean pH of 2 independent experiments in triplicate wells ( n = 6). Error bars represent standard deviation. Different letters represent statistically significant differences among treatments ( P
Figure Legend Snippet: Acidogenicity from Streptococcus mutans biofilms formed on enamel (A) and dentin (B), exposed to tested carbonated beverages. Plot shows pH of the culture medium. Each point in the plot represents mean pH of 2 independent experiments in triplicate wells ( n = 6). Error bars represent standard deviation. Different letters represent statistically significant differences among treatments ( P

Techniques Used: Standard Deviation

3) Product Images from "Assessment of in vivo versus in vitro biofilm formation of clinical methicillin-resistant Staphylococcus aureus isolates from endotracheal tubes"

Article Title: Assessment of in vivo versus in vitro biofilm formation of clinical methicillin-resistant Staphylococcus aureus isolates from endotracheal tubes

Journal: Scientific Reports

doi: 10.1038/s41598-018-30494-7

Biofilm production dynamics after extubation in 9 ETT-MRSA isolates under O 2 or 5% CO 2 Each color-bar represents the biofilm production of each ETT-MRSA isolate compared with the MRSA-in over days 1–8 after extubation. ( A ) Biofilm production dynamics under O 2 . Maximum biofilm production was on day 2. ( B ) Biofilm production dynamics under CO 2 . Maximum biofilm production was on day 1. Since the 5% CO 2 atmosphere better mimics the atmospheric conditions of mechanical ventilation. When ETT-MRSA are rapidly switched from the ETT environment to O 2 alone, they would need a day to adapt their metabolism to the new atmospheric conditions. Black points represent median biofilm production of the 9 isolates each day. Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Biofilm production dynamics after extubation in 9 ETT-MRSA isolates under O 2 or 5% CO 2 Each color-bar represents the biofilm production of each ETT-MRSA isolate compared with the MRSA-in over days 1–8 after extubation. ( A ) Biofilm production dynamics under O 2 . Maximum biofilm production was on day 2. ( B ) Biofilm production dynamics under CO 2 . Maximum biofilm production was on day 1. Since the 5% CO 2 atmosphere better mimics the atmospheric conditions of mechanical ventilation. When ETT-MRSA are rapidly switched from the ETT environment to O 2 alone, they would need a day to adapt their metabolism to the new atmospheric conditions. Black points represent median biofilm production of the 9 isolates each day. Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used:

Representative scanning electron microscopy of in vivo MRSA biofilm ( A ) Isolate 1 showing an in vivo detached biofilm at low magnification. Sometimes the sample processing for scanning electron microscopy released the biofilm cluster from the endotracheal tube surface. At higher magnification ( B ), cocci morphologies can be distinguished. The pig from which we obtained Isolate 1 was treated with vancomycin. ( C ) Isolate 45 (from a placebo treated pig) showing an in vivo biofilm attached to the endotracheal tube at low magnification. ( D ) at higher magnification, a cocci biofilm cluster was found (white arrow). Abbreviations: MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Representative scanning electron microscopy of in vivo MRSA biofilm ( A ) Isolate 1 showing an in vivo detached biofilm at low magnification. Sometimes the sample processing for scanning electron microscopy released the biofilm cluster from the endotracheal tube surface. At higher magnification ( B ), cocci morphologies can be distinguished. The pig from which we obtained Isolate 1 was treated with vancomycin. ( C ) Isolate 45 (from a placebo treated pig) showing an in vivo biofilm attached to the endotracheal tube at low magnification. ( D ) at higher magnification, a cocci biofilm cluster was found (white arrow). Abbreviations: MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used: Electron Microscopy, In Vivo

Biofilm production of 52 ETT-MRSA isolates compared with the MRSA-in under ambient air or ambient air with 5% CO 2. Each bar represents the biofilm production of each ETT-MRSA isolate versus the MRSA-in (black bar). ( A ) Biofilm production O 2 on day 2 after extubation (peak production); 50% of ETT-MRSA isolates increased biofilm production more than double that of MRSA-in. ( B ) Biofilm production under 5% CO 2 on day 1 after extubation (peak production); 40% of ETT-MRSA isolates increased more than twice MRSA-in biofilm production. The highest biofilm producers ETT-MRSA isolates (n = 9), in dark gray, under both O 2 and 5% CO 2 were selected to undergo the biofilm production dynamics post-extubation. Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Biofilm production of 52 ETT-MRSA isolates compared with the MRSA-in under ambient air or ambient air with 5% CO 2. Each bar represents the biofilm production of each ETT-MRSA isolate versus the MRSA-in (black bar). ( A ) Biofilm production O 2 on day 2 after extubation (peak production); 50% of ETT-MRSA isolates increased biofilm production more than double that of MRSA-in. ( B ) Biofilm production under 5% CO 2 on day 1 after extubation (peak production); 40% of ETT-MRSA isolates increased more than twice MRSA-in biofilm production. The highest biofilm producers ETT-MRSA isolates (n = 9), in dark gray, under both O 2 and 5% CO 2 were selected to undergo the biofilm production dynamics post-extubation. Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used:

Representative confocal laser scanning microscopy of in vivo MRSA biofilm Biofilm clusters (white arrows) were stained with the LIVE/DEAD BacLight kit (INVITROGEN, Barcelona, Spain). Viable bacteria (stained green by SYTO 9) are visible, but dead bacteria (stained red by propidium iodide) were infrequently detected. The nuclei and cytoplasm of eukaryotic cells from the pig were also stained nonspecifically by propidium iodide and SYTO 9 (large red and green blotches, respectively). ( A , B ) correspond to the in vivo biofilms of Isolate 39 and Isolate 45 obtained from pigs treated with vancomycin and placebo, respectively. Abbreviations: MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Representative confocal laser scanning microscopy of in vivo MRSA biofilm Biofilm clusters (white arrows) were stained with the LIVE/DEAD BacLight kit (INVITROGEN, Barcelona, Spain). Viable bacteria (stained green by SYTO 9) are visible, but dead bacteria (stained red by propidium iodide) were infrequently detected. The nuclei and cytoplasm of eukaryotic cells from the pig were also stained nonspecifically by propidium iodide and SYTO 9 (large red and green blotches, respectively). ( A , B ) correspond to the in vivo biofilms of Isolate 39 and Isolate 45 obtained from pigs treated with vancomycin and placebo, respectively. Abbreviations: MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used: Confocal Laser Scanning Microscopy, In Vivo, Staining

Effect of systemic antibiotic treatment on biofilm production in the 52 ETT-MRSA isolates under O 2 or 5% CO 2 conditions Median (interquartile range) values for biofilm production of the 52 ETT-MRSA compared with the MRSA-in under O 2 ( A ) and 5% CO 2 ( B ). Time of assessment: day of peak production. Biofilm production was not influenced by systemic treatment with placebo (n = 19), linezolid (n = 11), or vancomycin (n = 22) under either O 2 (1.96 [0.61–3.02], 2.30 [0.64–3.83], and 1.49 [0.63–2.67], respectively; p = 0.92) or 5% CO 2 (2.02 [1.16–2.34], 1.36 [0.48–2.75], and 1.09 [0.34–2.38], respectively; p = 0.62). Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Effect of systemic antibiotic treatment on biofilm production in the 52 ETT-MRSA isolates under O 2 or 5% CO 2 conditions Median (interquartile range) values for biofilm production of the 52 ETT-MRSA compared with the MRSA-in under O 2 ( A ) and 5% CO 2 ( B ). Time of assessment: day of peak production. Biofilm production was not influenced by systemic treatment with placebo (n = 19), linezolid (n = 11), or vancomycin (n = 22) under either O 2 (1.96 [0.61–3.02], 2.30 [0.64–3.83], and 1.49 [0.63–2.67], respectively; p = 0.92) or 5% CO 2 (2.02 [1.16–2.34], 1.36 [0.48–2.75], and 1.09 [0.34–2.38], respectively; p = 0.62). Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used:

4) Product Images from "Synergistic Interactions within a Multispecies Biofilm Enhance Individual Species Protection against Grazing by a Pelagic Protozoan"

Article Title: Synergistic Interactions within a Multispecies Biofilm Enhance Individual Species Protection against Grazing by a Pelagic Protozoan

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.02649

Change in viable cell numbers from the biofilm fractions of mono and multispecies cultures grazed with T. pyriformis after 24 and 96 h obtained from plating. The data points indicate the change in cell numbers of grazed biofilm fraction relative to the non-grazed biofilm fraction ± SEM obtained from two biological replicates.
Figure Legend Snippet: Change in viable cell numbers from the biofilm fractions of mono and multispecies cultures grazed with T. pyriformis after 24 and 96 h obtained from plating. The data points indicate the change in cell numbers of grazed biofilm fraction relative to the non-grazed biofilm fraction ± SEM obtained from two biological replicates.

Techniques Used:

Impact of grazing by T. pyriformis on the population dynamics of the individual bacterial species in the multispecies consortia as assessed by qPCR. Cell numbers of the individual bacterial members in (A) the multispecies biofilm fraction and (B) the multispecies planktonic fraction after 24 h of grazing.
Figure Legend Snippet: Impact of grazing by T. pyriformis on the population dynamics of the individual bacterial species in the multispecies consortia as assessed by qPCR. Cell numbers of the individual bacterial members in (A) the multispecies biofilm fraction and (B) the multispecies planktonic fraction after 24 h of grazing.

Techniques Used: Real-time Polymerase Chain Reaction

Grazing for 24 h by protozoa on the four-species mixed cultures. FISH based staining and confocal imaging shows the distribution of the different bacterial species in and around the protozoan cells. Applying the fluorescence filter channels, it is observed that X. retroflexus is abundantly present within the food vacuoles (indicated by the arrows) of T. pyriformis (A) . S. rhizophila (B) is detected to a lesser extent whereas M. oxydans (C) and P. amylolyticus (D) cells are not visibly present in the food vacuoles. (E,F) Depict the overlay and bright-field images, respectively. Panels (G,H) were included to phase out the dominating fluorescence signals from X. retroflexus and visualize the other bacterial members in the biofilm consortia around the ciliate.
Figure Legend Snippet: Grazing for 24 h by protozoa on the four-species mixed cultures. FISH based staining and confocal imaging shows the distribution of the different bacterial species in and around the protozoan cells. Applying the fluorescence filter channels, it is observed that X. retroflexus is abundantly present within the food vacuoles (indicated by the arrows) of T. pyriformis (A) . S. rhizophila (B) is detected to a lesser extent whereas M. oxydans (C) and P. amylolyticus (D) cells are not visibly present in the food vacuoles. (E,F) Depict the overlay and bright-field images, respectively. Panels (G,H) were included to phase out the dominating fluorescence signals from X. retroflexus and visualize the other bacterial members in the biofilm consortia around the ciliate.

Techniques Used: Fluorescence In Situ Hybridization, Staining, Imaging, Fluorescence

Biofilm forming index (BFI) of mono and mixed species cultures subject to Tetrahymena pyriformis (Tp) grazing and non-grazed cultures at 12, 24, and 96 h. The data points indicate the biofilm mean ± standard error of the mean (SEM) obtained from five biological replicates. ∗ P
Figure Legend Snippet: Biofilm forming index (BFI) of mono and mixed species cultures subject to Tetrahymena pyriformis (Tp) grazing and non-grazed cultures at 12, 24, and 96 h. The data points indicate the biofilm mean ± standard error of the mean (SEM) obtained from five biological replicates. ∗ P

Techniques Used:

Biofilm formation in the presence of T. pyriformis in three-species bacterial consortia. X. retroflexus is vital for biofilm development. Biofilm fold was calculated as the ratio of Abs 590 [(three – species biofilm cultured with grazer cells + SEM) – (three – species biofilm as control – SEM)] to Abs 590 (three – species biofilm cultured with grazer cells + SEM).
Figure Legend Snippet: Biofilm formation in the presence of T. pyriformis in three-species bacterial consortia. X. retroflexus is vital for biofilm development. Biofilm fold was calculated as the ratio of Abs 590 [(three – species biofilm cultured with grazer cells + SEM) – (three – species biofilm as control – SEM)] to Abs 590 (three – species biofilm cultured with grazer cells + SEM).

Techniques Used: Cell Culture

5) Product Images from "N-Acetyl-cysteine and Mechanisms Involved in Resolution of Chronic Wound Biofilm"

Article Title: N-Acetyl-cysteine and Mechanisms Involved in Resolution of Chronic Wound Biofilm

Journal: Journal of Diabetes Research

doi: 10.1155/2020/9589507

The effects of pH on the ability of NAC to dismantle the biofilm. (a) 24 h old biofilm was treated with either 20 or 10 mg/ml of NAC or HCL or acetic acid at different adjusted pH to 2.7, 3.1, 3.9, and 4.2, corresponding to the pH range of 20-3 mg/ml of NAC. When the pH of the NAC solution for both concentrations is below the pKa (3.24), biofilm was disturbed. However, when pH > pKa, the structure of the biofilm remained intact. Biofilm treated with HCL and acetic acid remained intact at ≥pH 3.1. When pH further decreased to pH 2.7, biofilm treated with both acids became fragile. (b) shows the pH of the NAC solution at the specific concentration of NAC. Three biological replicates were performed.
Figure Legend Snippet: The effects of pH on the ability of NAC to dismantle the biofilm. (a) 24 h old biofilm was treated with either 20 or 10 mg/ml of NAC or HCL or acetic acid at different adjusted pH to 2.7, 3.1, 3.9, and 4.2, corresponding to the pH range of 20-3 mg/ml of NAC. When the pH of the NAC solution for both concentrations is below the pKa (3.24), biofilm was disturbed. However, when pH > pKa, the structure of the biofilm remained intact. Biofilm treated with HCL and acetic acid remained intact at ≥pH 3.1. When pH further decreased to pH 2.7, biofilm treated with both acids became fragile. (b) shows the pH of the NAC solution at the specific concentration of NAC. Three biological replicates were performed.

Techniques Used: Concentration Assay

Detection of intracellular NAC using HPLC. 24 h-biofilm was treated with 20, 5 or 0 mg/ml NAC for 0 h or 4 h. After being collected and washed with PBS, cell pellets were sonicated to break the cell membrane and release intracellular NAC. The concentration of NAC was detected by an HPLC system equipped with a Phenomenex Luna C 18 column (150 × 4.6 mm; 5 μ m, 100 Å) and an UV/visible detector set at 214 nm. (a) Retention time for NAC was at 1.660 min. A standard curve was made with NAC concentrations ranging from 0 to 1 mg/ml, and from this curve, peak areas are used to determine the concentration of NAC in the experiment. (b) After 20 mg/ml NAC treatment for 0 and 4 h, the concentrations of intracellular NAC were significantly higher than both control and 5 mg/ml NAC. Biological duplicates from each treatment were used for the quantitative data.
Figure Legend Snippet: Detection of intracellular NAC using HPLC. 24 h-biofilm was treated with 20, 5 or 0 mg/ml NAC for 0 h or 4 h. After being collected and washed with PBS, cell pellets were sonicated to break the cell membrane and release intracellular NAC. The concentration of NAC was detected by an HPLC system equipped with a Phenomenex Luna C 18 column (150 × 4.6 mm; 5 μ m, 100 Å) and an UV/visible detector set at 214 nm. (a) Retention time for NAC was at 1.660 min. A standard curve was made with NAC concentrations ranging from 0 to 1 mg/ml, and from this curve, peak areas are used to determine the concentration of NAC in the experiment. (b) After 20 mg/ml NAC treatment for 0 and 4 h, the concentrations of intracellular NAC were significantly higher than both control and 5 mg/ml NAC. Biological duplicates from each treatment were used for the quantitative data.

Techniques Used: High Performance Liquid Chromatography, Sonication, Concentration Assay

The effect of NAC on biofilm development and dismantling. (a) Microbiome taken from the chronic wounds and grown in 96-well microtiter plates. Concentrations of NAC ranging from 3 to 20 mg/ml were applied to the cultures at the times indicated (0, 6, 12, and 24 h). The pictures shown were taken at 48 h after initiation of the culture. When ≥4 mg/ml NAC were applied to the culture at 0 h, the biofilm never developed. When ≥10 mg/ml of NAC was applied at 6, 12, and 24 h after culture initiation, biofilm formation was visibly altered or became fragile and disrupted. (b) At 48 h, biofilm was stained with crystal violet (CV) and the absorbance at 590 nm was measured to quantify biofilm biomass. At 0 h, 5 mg/ml application, 5 mg/ml NAC significantly reduced the appearance of biofilm, and when applied at 6 and 12 h, it was able to decrease biofilm production but not when applied at 24 h. ≥10 mg/ml NAC or more significantly diminish biofilm formation. Three technical repeats with standard deviation as error bars for quantitative data. (c) The dose-dependent effect of NAC on 24 h old biofilm cultured in 35 mm petri dishes was recorded over time. 20 mg/ml of NAC dismantled biofilm by 24 h; 10 and 7.5 mg/ml of NAC were able to fully disrupt biofilm at 30 and 54 h, respectively. 5 mg/ml NAC did not cause changes in existing biofilm.
Figure Legend Snippet: The effect of NAC on biofilm development and dismantling. (a) Microbiome taken from the chronic wounds and grown in 96-well microtiter plates. Concentrations of NAC ranging from 3 to 20 mg/ml were applied to the cultures at the times indicated (0, 6, 12, and 24 h). The pictures shown were taken at 48 h after initiation of the culture. When ≥4 mg/ml NAC were applied to the culture at 0 h, the biofilm never developed. When ≥10 mg/ml of NAC was applied at 6, 12, and 24 h after culture initiation, biofilm formation was visibly altered or became fragile and disrupted. (b) At 48 h, biofilm was stained with crystal violet (CV) and the absorbance at 590 nm was measured to quantify biofilm biomass. At 0 h, 5 mg/ml application, 5 mg/ml NAC significantly reduced the appearance of biofilm, and when applied at 6 and 12 h, it was able to decrease biofilm production but not when applied at 24 h. ≥10 mg/ml NAC or more significantly diminish biofilm formation. Three technical repeats with standard deviation as error bars for quantitative data. (c) The dose-dependent effect of NAC on 24 h old biofilm cultured in 35 mm petri dishes was recorded over time. 20 mg/ml of NAC dismantled biofilm by 24 h; 10 and 7.5 mg/ml of NAC were able to fully disrupt biofilm at 30 and 54 h, respectively. 5 mg/ml NAC did not cause changes in existing biofilm.

Techniques Used: Staining, Standard Deviation, Cell Culture

The effects of NAC and similar molecules in dismantling biofilm. (a) Structures of molecules similar to NAC used in this experiment: N -acetyl-cysteine amide (NACA), N -acetyl-serine (NAS), cysteine (Cys), and glutathione (GTH). (b) Chronic wound microbiome biofilm formation after the treatment at time 0 h with the different molecules with concentration ranging from 0 to 20 mg/ml. > 4 mg/ml of NAC, NAS, and GTH interfered with chronic wound microbiome cell growth. However, the same amount of NACA and cysteine did not affect cell growth. (c) Quantification of the biofilm formation from (b) using crystal violet (CV) staining. Compared to LB media as the control, ≥4 mg/ml NAC, NAS, and GTH were able to stop the formation of biofilm. ≥4 mg/ml NACA did not affect cell growth in (b), yet significantly reduced biofilm. The application of 3-20 mg/ml Cys to chronic wound microbiome culture did not increase or reduce biofilm formation. (d) 24 h chronic wound microbiome biofilm was treated with various molecules and the effects recorded at 48 h. Biofilm was visually dismantled when treated with ≥10 mg/ml NAC and 20 mg/ml NAS. NACA, cysteine, and GTH did not seem to affect the biofilm. (e) Quantification of existing biofilm formation from (d) using CV staining. ≥7.5 mg/ml NAC and ≥10 mg/ml NAS can significantly decrease existing biofilm. Technical triplicates for each treatment for both cultured biofilm and quantitative data.
Figure Legend Snippet: The effects of NAC and similar molecules in dismantling biofilm. (a) Structures of molecules similar to NAC used in this experiment: N -acetyl-cysteine amide (NACA), N -acetyl-serine (NAS), cysteine (Cys), and glutathione (GTH). (b) Chronic wound microbiome biofilm formation after the treatment at time 0 h with the different molecules with concentration ranging from 0 to 20 mg/ml. > 4 mg/ml of NAC, NAS, and GTH interfered with chronic wound microbiome cell growth. However, the same amount of NACA and cysteine did not affect cell growth. (c) Quantification of the biofilm formation from (b) using crystal violet (CV) staining. Compared to LB media as the control, ≥4 mg/ml NAC, NAS, and GTH were able to stop the formation of biofilm. ≥4 mg/ml NACA did not affect cell growth in (b), yet significantly reduced biofilm. The application of 3-20 mg/ml Cys to chronic wound microbiome culture did not increase or reduce biofilm formation. (d) 24 h chronic wound microbiome biofilm was treated with various molecules and the effects recorded at 48 h. Biofilm was visually dismantled when treated with ≥10 mg/ml NAC and 20 mg/ml NAS. NACA, cysteine, and GTH did not seem to affect the biofilm. (e) Quantification of existing biofilm formation from (d) using CV staining. ≥7.5 mg/ml NAC and ≥10 mg/ml NAS can significantly decrease existing biofilm. Technical triplicates for each treatment for both cultured biofilm and quantitative data.

Techniques Used: Concentration Assay, Staining, Cell Culture

The effect of NAC on the protein and DNA of the EPS. 24 h chronic wound microbiome biofilm was treated with NAC at different concentrations for 24 h. The same amount of biofilm inoculum was extracted and analyzed on 12% SDS-PAGE gels. (a) Whole protein extract. (b) Soluble protein component (EPS fraction). In the presence of 20 mg/ml NAC, there were significantly less proteins both in the whole extract and in the soluble fraction. With 10 mg/ml NAC, the protein in the whole extract was not different from that of the control whereas the protein in the supernatant was significantly decreased. NAC treatment with less than of 7.5 mg/ml did not alter either the whole extract or the soluble proteins except for those above ~60 kDa. (c) DNA components in the biofilm matrix were analyzed using 1% agarose gels. A prominent band was seen at 45.8 kb with 5, 7.5, and 10 mg/ml NAC treatments. Treatment with 20 mg/ml of NAC reduced the amount of DNA present in that band, and no other DNA bands were seen. The pattern of DNA in the EPS with 5 mg/ml NAC treatment was similar to that of the control and showed not only the 45.8 kb band but also other smaller bands:
Figure Legend Snippet: The effect of NAC on the protein and DNA of the EPS. 24 h chronic wound microbiome biofilm was treated with NAC at different concentrations for 24 h. The same amount of biofilm inoculum was extracted and analyzed on 12% SDS-PAGE gels. (a) Whole protein extract. (b) Soluble protein component (EPS fraction). In the presence of 20 mg/ml NAC, there were significantly less proteins both in the whole extract and in the soluble fraction. With 10 mg/ml NAC, the protein in the whole extract was not different from that of the control whereas the protein in the supernatant was significantly decreased. NAC treatment with less than of 7.5 mg/ml did not alter either the whole extract or the soluble proteins except for those above ~60 kDa. (c) DNA components in the biofilm matrix were analyzed using 1% agarose gels. A prominent band was seen at 45.8 kb with 5, 7.5, and 10 mg/ml NAC treatments. Treatment with 20 mg/ml of NAC reduced the amount of DNA present in that band, and no other DNA bands were seen. The pattern of DNA in the EPS with 5 mg/ml NAC treatment was similar to that of the control and showed not only the 45.8 kb band but also other smaller bands:

Techniques Used: SDS Page

Fluorescence staining and confocal laser scanning microscopy analysis of the biofilm: DNA (green), proteins (red), and polysaccharides (blue) were visualized by staining with specific stains as described in the Methods and Materials section. Pictures in (a) show that without the application of NAC, extracellular DNA molecules appear as strings in the matrix (arrowheads). The proteins are primarily found either in the bacteria which were planktonic or in aggregates. The carbohydrates are mostly in association with the planktonic bacteria and the bacterial aggregates. Pictures in (b) show that treatment with 10 mg/ml NAC resulted in virtually no bacteria present and the EPS was found to be mostly gone. Biological duplicates for each treatment were used for the images.
Figure Legend Snippet: Fluorescence staining and confocal laser scanning microscopy analysis of the biofilm: DNA (green), proteins (red), and polysaccharides (blue) were visualized by staining with specific stains as described in the Methods and Materials section. Pictures in (a) show that without the application of NAC, extracellular DNA molecules appear as strings in the matrix (arrowheads). The proteins are primarily found either in the bacteria which were planktonic or in aggregates. The carbohydrates are mostly in association with the planktonic bacteria and the bacterial aggregates. Pictures in (b) show that treatment with 10 mg/ml NAC resulted in virtually no bacteria present and the EPS was found to be mostly gone. Biological duplicates for each treatment were used for the images.

Techniques Used: Fluorescence, Staining, Confocal Laser Scanning Microscopy

The effect of NAC on the growth of bacteria contained in biofilm. We used the culture shown in Figure 2(c) to perform this experiment. (a) NAC was applied to the culture from one side of the culture dish marked with an arrowhead. The other side of the plate where NAC was not applied is marked by a star. (b) Efficiency of NAC in penetrating the biofilm and interfering with bacterial cell growth. After 0, 1.5 h, 4 h, and 9 h NAC treatments, cells from the side where NAC was applied (arrowhead) and away from the site of application (star) were subcultured in fresh LB in a microtiter plate. 20 mg/ml NAC were able to penetrate the culture and interfere with cell growth within 1.5 h, and no cell growth was detected after 4 h treatment. 10 and 7.5 mg/ml NAC were able to penetrate the culture and completely inhibit further cell growth after 9 h post application. 5 mg/ml NAC could not inhibit cell growth. Technical duplicates were used for cell viabilities after being treated with different concentrations of NAC.
Figure Legend Snippet: The effect of NAC on the growth of bacteria contained in biofilm. We used the culture shown in Figure 2(c) to perform this experiment. (a) NAC was applied to the culture from one side of the culture dish marked with an arrowhead. The other side of the plate where NAC was not applied is marked by a star. (b) Efficiency of NAC in penetrating the biofilm and interfering with bacterial cell growth. After 0, 1.5 h, 4 h, and 9 h NAC treatments, cells from the side where NAC was applied (arrowhead) and away from the site of application (star) were subcultured in fresh LB in a microtiter plate. 20 mg/ml NAC were able to penetrate the culture and interfere with cell growth within 1.5 h, and no cell growth was detected after 4 h treatment. 10 and 7.5 mg/ml NAC were able to penetrate the culture and completely inhibit further cell growth after 9 h post application. 5 mg/ml NAC could not inhibit cell growth. Technical duplicates were used for cell viabilities after being treated with different concentrations of NAC.

Techniques Used:

Chronic wound microbiome rich in P. aeruginosa formed strong biofilm in vitro . Microbiome rich in P. aeruginosa was collected from chronic wounds and cultured in vitro overnight. Similar cultures were performed with microbiome collected from nonchronic wound. (a) Bacterial species sequencing data using the 16S ITS region as the probe. In this particular animal, the wound became colonized with P. aeruginosa as early as day 3 postwounding and induction of chronicity. (b) Microbiome cultures in 96-well microtiter plates. Top view, side view, and side view with crystal violet staining. Microbiome from chronic wounds formed thick opaque biofilm on the air-liquid interface that can be stained with crystal violet (long arrow). Microbiome from nonchronic wounds did not form biofilm. Technical replicas of three wells of each condition. (c) Likewise, when cultured in tubes, microbiome from nonchronic wounds did not stain with crystal violet (left 2 tubes) whereas that from chronic wounds was stained with crystal violet (right 2 tubes).
Figure Legend Snippet: Chronic wound microbiome rich in P. aeruginosa formed strong biofilm in vitro . Microbiome rich in P. aeruginosa was collected from chronic wounds and cultured in vitro overnight. Similar cultures were performed with microbiome collected from nonchronic wound. (a) Bacterial species sequencing data using the 16S ITS region as the probe. In this particular animal, the wound became colonized with P. aeruginosa as early as day 3 postwounding and induction of chronicity. (b) Microbiome cultures in 96-well microtiter plates. Top view, side view, and side view with crystal violet staining. Microbiome from chronic wounds formed thick opaque biofilm on the air-liquid interface that can be stained with crystal violet (long arrow). Microbiome from nonchronic wounds did not form biofilm. Technical replicas of three wells of each condition. (c) Likewise, when cultured in tubes, microbiome from nonchronic wounds did not stain with crystal violet (left 2 tubes) whereas that from chronic wounds was stained with crystal violet (right 2 tubes).

Techniques Used: In Vitro, Cell Culture, Sequencing, Staining

The effects of NAC on bacterial oxidative stress and protein synthesis. (a) NAD + /NADH ratio for the 24 h biofilm treated with NAC. The ratio decreases with the increase of NAC concentration indicating that NAC treatment causes an increase in oxidative stress in the cytosol of the bacteria. (b) 35 S labeling of proteins shows that NAC at 20 mg/ml inhibits protein synthesis whereas at 5 mg/ml does not.
Figure Legend Snippet: The effects of NAC on bacterial oxidative stress and protein synthesis. (a) NAD + /NADH ratio for the 24 h biofilm treated with NAC. The ratio decreases with the increase of NAC concentration indicating that NAC treatment causes an increase in oxidative stress in the cytosol of the bacteria. (b) 35 S labeling of proteins shows that NAC at 20 mg/ml inhibits protein synthesis whereas at 5 mg/ml does not.

Techniques Used: Concentration Assay, Labeling

6) Product Images from "Preparation of PLGA-chitosan based nanocarriers for enhancing antibacterial effect of ciprofloxacin in root canal infection"

Article Title: Preparation of PLGA-chitosan based nanocarriers for enhancing antibacterial effect of ciprofloxacin in root canal infection

Journal: Drug Delivery

doi: 10.1080/10717544.2019.1701140

Statistical analysis of biofilm inhibition assay (mean ± SD, n = 3).
Figure Legend Snippet: Statistical analysis of biofilm inhibition assay (mean ± SD, n = 3).

Techniques Used: Inhibition

7) Product Images from "Lipopeptide biosurfactant viscosin enhances dispersal of Pseudomonas fluorescens SBW25 biofilms"

Article Title: Lipopeptide biosurfactant viscosin enhances dispersal of Pseudomonas fluorescens SBW25 biofilms

Journal: Microbiology

doi: 10.1099/mic.0.000191

Expression of the viscA gene per biomass unit in biofilms (blue diamonds) and planktonic cells (red squares) in the initial state of biofilms grown in microtitre wells. Data represent mean ± sd for a representative experiment with five replicates. *Significant difference between biofilm and planktonic cells at each time point ( P
Figure Legend Snippet: Expression of the viscA gene per biomass unit in biofilms (blue diamonds) and planktonic cells (red squares) in the initial state of biofilms grown in microtitre wells. Data represent mean ± sd for a representative experiment with five replicates. *Significant difference between biofilm and planktonic cells at each time point ( P

Techniques Used: Expressing

Biofilms of P. fluorescens SBW25 (WT) and SBW25Δ viscA ( viscA mutant) were cultivated in flow chamber systems with AB minimal medium with citrate for 1 day, upon which a shift was made to AB minimal medium with no carbon source. CLSM micrographs were acquired immediately before the shift (dark bars) and 3 h after the shift (light bars), and the biomass was quantified using comstat . Data represent mean ± sd of 18 images taken from random places in three independent flow chambers. *Significant difference between pairs of values ( P
Figure Legend Snippet: Biofilms of P. fluorescens SBW25 (WT) and SBW25Δ viscA ( viscA mutant) were cultivated in flow chamber systems with AB minimal medium with citrate for 1 day, upon which a shift was made to AB minimal medium with no carbon source. CLSM micrographs were acquired immediately before the shift (dark bars) and 3 h after the shift (light bars), and the biomass was quantified using comstat . Data represent mean ± sd of 18 images taken from random places in three independent flow chambers. *Significant difference between pairs of values ( P

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

CLSM micrographs of biofilms of P. fluorescens (a, b) SBW25 Tn 7 :: gfp 2 and (c, d) SBW25 Tn 7 :: gfp2 Δ viscA . The strains were cultivated in flow chamber systems for 1 day with AB minimal medium with citrate, upon which a shift was made to AB minimal medium with no carbon source. Micrographs were acquired immediately before the shift (a, c) and 3 h after the shift (b, d). Bars, 20 μm.
Figure Legend Snippet: CLSM micrographs of biofilms of P. fluorescens (a, b) SBW25 Tn 7 :: gfp 2 and (c, d) SBW25 Tn 7 :: gfp2 Δ viscA . The strains were cultivated in flow chamber systems for 1 day with AB minimal medium with citrate, upon which a shift was made to AB minimal medium with no carbon source. Micrographs were acquired immediately before the shift (a, c) and 3 h after the shift (b, d). Bars, 20 μm.

Techniques Used: Confocal Laser Scanning Microscopy, Flow Cytometry

Mature biofilms of P. fluorescens SBW25Δ viscA complemented with purified viscosin at concentrations of 0 and 50 μg ml − 1 . Biofilms were quantified after 9.5 h of incubation with and without viscosin by the crystal violet assay. Data represent mean ± sd for a representative dataset including eight replicates. *Significant difference ( P
Figure Legend Snippet: Mature biofilms of P. fluorescens SBW25Δ viscA complemented with purified viscosin at concentrations of 0 and 50 μg ml − 1 . Biofilms were quantified after 9.5 h of incubation with and without viscosin by the crystal violet assay. Data represent mean ± sd for a representative dataset including eight replicates. *Significant difference ( P

Techniques Used: Purification, Incubation, Crystal Violet Assay

CLSM micrographs of biofilms formed by P. fluorescens SBW25 Tn 7 :: gfp 2pSEVA237R_P viscA. The images were acquired 3 h after a shift to AB minimal medium with no carbon source. Artificial colours have been assigned in these images to the different CLSM signals; green denotes cells expressing the GFP marker; red denotes cells expressing the viscA -mCherry reporter. (a) GFP and mCherry. (b) mCherry only. Arrows indicate selected microcolonies that are only green and not red.
Figure Legend Snippet: CLSM micrographs of biofilms formed by P. fluorescens SBW25 Tn 7 :: gfp 2pSEVA237R_P viscA. The images were acquired 3 h after a shift to AB minimal medium with no carbon source. Artificial colours have been assigned in these images to the different CLSM signals; green denotes cells expressing the GFP marker; red denotes cells expressing the viscA -mCherry reporter. (a) GFP and mCherry. (b) mCherry only. Arrows indicate selected microcolonies that are only green and not red.

Techniques Used: Confocal Laser Scanning Microscopy, Expressing, Marker

CLSM micrographs of biofilms formed by P. fluorescens (a) SBW25 Tn 7 :: gfp2 and (b) SBW25 Tn 7 :: gfp2 Δ viscA grown in AB minimal medium with citrate in flow cells for 2 days. Bars, 20 μm.
Figure Legend Snippet: CLSM micrographs of biofilms formed by P. fluorescens (a) SBW25 Tn 7 :: gfp2 and (b) SBW25 Tn 7 :: gfp2 Δ viscA grown in AB minimal medium with citrate in flow cells for 2 days. Bars, 20 μm.

Techniques Used: Confocal Laser Scanning Microscopy, Flow Cytometry

Biofilm formation of P. fluorescens SBW25 (dark bars) and P. fluorescens SBW25Δ viscA (light bars) in AB minimal medium with citrate. The biofilm was quantified from 7.5 to 17.5 h using the crystal violet assay. The A 590 value represents crystal violet-stained biofilm attached to the walls of the microtitre wells and is an indirect measure of the biofilm formed. Data represent mean ± sd for a representative experiment with eight replicates. *Significant difference between strains at each time point ( P
Figure Legend Snippet: Biofilm formation of P. fluorescens SBW25 (dark bars) and P. fluorescens SBW25Δ viscA (light bars) in AB minimal medium with citrate. The biofilm was quantified from 7.5 to 17.5 h using the crystal violet assay. The A 590 value represents crystal violet-stained biofilm attached to the walls of the microtitre wells and is an indirect measure of the biofilm formed. Data represent mean ± sd for a representative experiment with eight replicates. *Significant difference between strains at each time point ( P

Techniques Used: Crystal Violet Assay, Staining

8) Product Images from "Molecular Characteristics of Staphylococcus aureus Causing Bovine Mastitis between 2014 and 2015"

Article Title: Molecular Characteristics of Staphylococcus aureus Causing Bovine Mastitis between 2014 and 2015

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2017.00127

Biofilm formation and red blood cells lysis. (A) Semi-quantitative biofilm analysis of STs with more than 10 isolates. *** P
Figure Legend Snippet: Biofilm formation and red blood cells lysis. (A) Semi-quantitative biofilm analysis of STs with more than 10 isolates. *** P

Techniques Used: Lysis

9) Product Images from "Disease-induced assemblage of a plant-beneficial bacterial consortium"

Article Title: Disease-induced assemblage of a plant-beneficial bacterial consortium

Journal: The ISME Journal

doi: 10.1038/s41396-018-0093-1

Synergistic interactions between recruited Xanthomonas , Stenotrophomonas , and Microbacterium spp. strains. a Boxplot of biofilm formation by single Xanthomonas sp. WCS2014-23 (X; eOTU 106), Stenotrophomonas sp. WCS2014-113 (S; eOTU 97) and Microbacterium sp. WCS2014-259 (M; eOTU 107) or the double (XS, XM, and SM) and triple combinations (XSM) thereof in Nunc-TSP lid plates. After 24 h of incubation, the biofilm formation was quantified by staining with crystal violet. b Attraction between colonies of X, S, and M grown at increasing proximity on King’s medium B agar
Figure Legend Snippet: Synergistic interactions between recruited Xanthomonas , Stenotrophomonas , and Microbacterium spp. strains. a Boxplot of biofilm formation by single Xanthomonas sp. WCS2014-23 (X; eOTU 106), Stenotrophomonas sp. WCS2014-113 (S; eOTU 97) and Microbacterium sp. WCS2014-259 (M; eOTU 107) or the double (XS, XM, and SM) and triple combinations (XSM) thereof in Nunc-TSP lid plates. After 24 h of incubation, the biofilm formation was quantified by staining with crystal violet. b Attraction between colonies of X, S, and M grown at increasing proximity on King’s medium B agar

Techniques Used: Incubation, Staining

10) Product Images from "Disease-induced assemblage of a plant-beneficial bacterial consortium"

Article Title: Disease-induced assemblage of a plant-beneficial bacterial consortium

Journal: The ISME Journal

doi: 10.1038/s41396-018-0093-1

Synergistic interactions between recruited Xanthomonas , Stenotrophomonas , and Microbacterium spp. strains. a Boxplot of biofilm formation by single Xanthomonas sp. WCS2014-23 (X; eOTU 106), Stenotrophomonas sp. WCS2014-113 (S; eOTU 97) and Microbacterium sp. WCS2014-259 (M; eOTU 107) or the double (XS, XM, and SM) and triple combinations (XSM) thereof in Nunc-TSP lid plates. After 24 h of incubation, the biofilm formation was quantified by staining with crystal violet. b Attraction between colonies of X, S, and M grown at increasing proximity on King’s medium B agar
Figure Legend Snippet: Synergistic interactions between recruited Xanthomonas , Stenotrophomonas , and Microbacterium spp. strains. a Boxplot of biofilm formation by single Xanthomonas sp. WCS2014-23 (X; eOTU 106), Stenotrophomonas sp. WCS2014-113 (S; eOTU 97) and Microbacterium sp. WCS2014-259 (M; eOTU 107) or the double (XS, XM, and SM) and triple combinations (XSM) thereof in Nunc-TSP lid plates. After 24 h of incubation, the biofilm formation was quantified by staining with crystal violet. b Attraction between colonies of X, S, and M grown at increasing proximity on King’s medium B agar

Techniques Used: Incubation, Staining

11) Product Images from "Bacterial Cyclic AMP-Phosphodiesterase Activity Coordinates Biofilm Formation"

Article Title: Bacterial Cyclic AMP-Phosphodiesterase Activity Coordinates Biofilm Formation

Journal: PLoS ONE

doi: 10.1371/journal.pone.0071267

CpdS mediates S. marcescens biofilm formation. A–B . Quantification of crystal violet stained biofilms from the sides of glass test tubes grown under high-sheer conditions. Vector = pMQ131; p cpdS = pMQ171; pN94A = pMQ176. The asterisk indicates statistical difference from wild-type levels (n = 9, p
Figure Legend Snippet: CpdS mediates S. marcescens biofilm formation. A–B . Quantification of crystal violet stained biofilms from the sides of glass test tubes grown under high-sheer conditions. Vector = pMQ131; p cpdS = pMQ171; pN94A = pMQ176. The asterisk indicates statistical difference from wild-type levels (n = 9, p

Techniques Used: Staining, Plasmid Preparation

Role of cAMP-phosphodiesterase activity in biofilm formation and identification of a cAMP-phosphodiesterase gene in the S. marcescens genome. A. Model for cAMP metabolism and inhibitory effect on biofilm production. Adenylate cyclase (AC) catalyzes synthesis of cAMP from ATP, whereas cyclic-AMP phosphodiesterase (cAMP-PDE) catalyzes hydrolysis of cAMP to 5′-AMP. B. Crystal violet stained biofilms on the side of test tubes formed under high-sheer conditions. Shown is a wild-type S. marcescens strain with either the empty vector or the vector with a wild-type copy of the E. coli cAMP-PDE gene, cpdA . C. Genomic context of the S. marcescens cpdS gene, a candidate cAMP-PDE gene.
Figure Legend Snippet: Role of cAMP-phosphodiesterase activity in biofilm formation and identification of a cAMP-phosphodiesterase gene in the S. marcescens genome. A. Model for cAMP metabolism and inhibitory effect on biofilm production. Adenylate cyclase (AC) catalyzes synthesis of cAMP from ATP, whereas cyclic-AMP phosphodiesterase (cAMP-PDE) catalyzes hydrolysis of cAMP to 5′-AMP. B. Crystal violet stained biofilms on the side of test tubes formed under high-sheer conditions. Shown is a wild-type S. marcescens strain with either the empty vector or the vector with a wild-type copy of the E. coli cAMP-PDE gene, cpdA . C. Genomic context of the S. marcescens cpdS gene, a candidate cAMP-PDE gene.

Techniques Used: Activity Assay, Staining, Plasmid Preparation

12) Product Images from "The antibacterial effect of non-thermal atmospheric pressure plasma treatment of titanium surfaces according to the bacterial wall structure"

Article Title: The antibacterial effect of non-thermal atmospheric pressure plasma treatment of titanium surfaces according to the bacterial wall structure

Journal: Scientific Reports

doi: 10.1038/s41598-019-39414-9

Changes in rate of biofilm formation ability by four bacteria species. *No differences among the NTAPPJ-treated time groups (p > 0.05). The same lowercase letter indicates no significant difference among the bacteria species groups (p > 0.05).
Figure Legend Snippet: Changes in rate of biofilm formation ability by four bacteria species. *No differences among the NTAPPJ-treated time groups (p > 0.05). The same lowercase letter indicates no significant difference among the bacteria species groups (p > 0.05).

Techniques Used:

13) Product Images from "CRISPR-Cas9 modified bacteriophage for treatment of Staphylococcus aureus induced osteomyelitis and soft tissue infection"

Article Title: CRISPR-Cas9 modified bacteriophage for treatment of Staphylococcus aureus induced osteomyelitis and soft tissue infection

Journal: PLoS ONE

doi: 10.1371/journal.pone.0220421

Bacteria on ex vivo orthopedic screws and bone. (A-B) Gram positive cocci, and what appears to be biofilm, was evident on the distal portion (A) and between threads (B) of an ex vivo orthopedic screw excised at day 7. (C-D). Separate fragments adjacent to the defect/screw site from a representative untreated (empty alginate) control femur collected at day 8 revealed dispersed Gram positive cocci.
Figure Legend Snippet: Bacteria on ex vivo orthopedic screws and bone. (A-B) Gram positive cocci, and what appears to be biofilm, was evident on the distal portion (A) and between threads (B) of an ex vivo orthopedic screw excised at day 7. (C-D). Separate fragments adjacent to the defect/screw site from a representative untreated (empty alginate) control femur collected at day 8 revealed dispersed Gram positive cocci.

Techniques Used: Ex Vivo

14) Product Images from "Bacterial fight-and-flight responses enhance virulence in a polymicrobial infection"

Article Title: Bacterial fight-and-flight responses enhance virulence in a polymicrobial infection

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1400586111

dspB mediates dispersal in response to H 2 O 2 . ( A ) Quantitative microtiter dish assay for measuring biofilm dispersal. (1) Aa is incubated statically under anoxic conditions to promote attachment to the bottom of microtiter dish wells, (2) biofilms are incubated under oxic shaking conditions either with (+H 2 O 2 ) or without H 2 O 2 addition, and (3) biofilm mass is quantified using crystal violet. ( B ) Crystal violet incorporated into biofilms was solubilized and quantified by measuring absorbance. The y axis represents the absorbance reading for H 2 O 2 -treated biofilms divided by the reading for untreated biofilms. A value less than one thus indicates that dispersal occurred, whereas a value close to one indicates that dispersal was absent. Error bars represent SEM ( n = 4). * P
Figure Legend Snippet: dspB mediates dispersal in response to H 2 O 2 . ( A ) Quantitative microtiter dish assay for measuring biofilm dispersal. (1) Aa is incubated statically under anoxic conditions to promote attachment to the bottom of microtiter dish wells, (2) biofilms are incubated under oxic shaking conditions either with (+H 2 O 2 ) or without H 2 O 2 addition, and (3) biofilm mass is quantified using crystal violet. ( B ) Crystal violet incorporated into biofilms was solubilized and quantified by measuring absorbance. The y axis represents the absorbance reading for H 2 O 2 -treated biofilms divided by the reading for untreated biofilms. A value less than one thus indicates that dispersal occurred, whereas a value close to one indicates that dispersal was absent. Error bars represent SEM ( n = 4). * P

Techniques Used: Incubation

dspB is induced by oxygen. ( A ) β-galactosidase activity was quantified for anoxically grown colony biofilms of Aa strain 624 that were transferred to media alone or media supplemented with exogenous catalase (10,000 U mL −1 ) and incubated under anoxic or oxic conditions for 6 h. Fold difference in dspB expression ( y axis) was determined by dividing the expression value for each condition by the corresponding anoxic value. The dashed line represents anoxic expression (normalized to one within each replicate). Error bars represent SEM ( n = 3). * P
Figure Legend Snippet: dspB is induced by oxygen. ( A ) β-galactosidase activity was quantified for anoxically grown colony biofilms of Aa strain 624 that were transferred to media alone or media supplemented with exogenous catalase (10,000 U mL −1 ) and incubated under anoxic or oxic conditions for 6 h. Fold difference in dspB expression ( y axis) was determined by dividing the expression value for each condition by the corresponding anoxic value. The dashed line represents anoxic expression (normalized to one within each replicate). Error bars represent SEM ( n = 3). * P

Techniques Used: Activity Assay, Incubation, Expressing

dspB transcription is enhanced during coculture with Sg . Anoxically grown mono- and coculture colony biofilms were transferred to fresh media and incubated for 1 h under oxic or anoxic conditions before measuring β-gal activity. Coculture biofilms were formed by thoroughly mixing equal volumes of exponential-phase cultures of either Aa and Sg or Aa and Sg spxB − that were each adjusted to an OD = 1. Fold difference in dspB expression ( y axis) was calculated by dividing the expression value for each condition by the anoxic monoculture value (normalized to one). Error bars represent SEM. The oxic Aa + Sg spxB − coculture value was not significantly larger than the oxic Aa monoculture value ( P > 0.13 by a two-tailed t test). ** P
Figure Legend Snippet: dspB transcription is enhanced during coculture with Sg . Anoxically grown mono- and coculture colony biofilms were transferred to fresh media and incubated for 1 h under oxic or anoxic conditions before measuring β-gal activity. Coculture biofilms were formed by thoroughly mixing equal volumes of exponential-phase cultures of either Aa and Sg or Aa and Sg spxB − that were each adjusted to an OD = 1. Fold difference in dspB expression ( y axis) was calculated by dividing the expression value for each condition by the anoxic monoculture value (normalized to one). Error bars represent SEM. The oxic Aa + Sg spxB − coculture value was not significantly larger than the oxic Aa monoculture value ( P > 0.13 by a two-tailed t test). ** P

Techniques Used: Incubation, Activity Assay, Expressing, Two Tailed Test

15) Product Images from "Enterococcal Surface Protein, Esp, Enhances Biofilm Formation by Enterococcus faecalis"

Article Title: Enterococcal Surface Protein, Esp, Enhances Biofilm Formation by Enterococcus faecalis

Journal: Infection and Immunity

doi: 10.1128/IAI.72.10.6032-6039.2004

Biofilm formation by Esp-negative FA2-2(pAT28), Esp-positive FA2-2(pESPF), GelE-positive FA2-2(pCUGel), and Esp- and GelE-positive FA2-2(pCUGel, pESPF) on 96-well polystyrene plates as quantified by crystal violet staining. The error bars represent mean
Figure Legend Snippet: Biofilm formation by Esp-negative FA2-2(pAT28), Esp-positive FA2-2(pESPF), GelE-positive FA2-2(pCUGel), and Esp- and GelE-positive FA2-2(pCUGel, pESPF) on 96-well polystyrene plates as quantified by crystal violet staining. The error bars represent mean

Techniques Used: End-sequence Profiling, Staining

Biofilm formation by Esp-positive and Esp-negative strains at different glucose concentrations. Biofilm formation was quantified by crystal violet staining, followed by back extraction of bound crystal violet into an ethanol-acetone mixture. The y axis
Figure Legend Snippet: Biofilm formation by Esp-positive and Esp-negative strains at different glucose concentrations. Biofilm formation was quantified by crystal violet staining, followed by back extraction of bound crystal violet into an ethanol-acetone mixture. The y axis

Techniques Used: End-sequence Profiling, Staining

Biofilm densities (□) and cell surface hydrophobicities (•) of Esp-positive and Esp-negative FA2-2 and OG1RF strain pairs. The bacteria were cultured in TSB containing 0.75% glucose, and the cell surface hydrophobicities were measured
Figure Legend Snippet: Biofilm densities (□) and cell surface hydrophobicities (•) of Esp-positive and Esp-negative FA2-2 and OG1RF strain pairs. The bacteria were cultured in TSB containing 0.75% glucose, and the cell surface hydrophobicities were measured

Techniques Used: Cell Surface Hydrophobicity, End-sequence Profiling, Cell Culture

Esp-mediated biofilm formation on polyvinyl chloride and polypropylene substrates. The ability of Esp-positive and Esp-negative strains to form biofilms on medically relevant surfaces such as polyvinyl chloride and polypropylene was assessed by crystal
Figure Legend Snippet: Esp-mediated biofilm formation on polyvinyl chloride and polypropylene substrates. The ability of Esp-positive and Esp-negative strains to form biofilms on medically relevant surfaces such as polyvinyl chloride and polypropylene was assessed by crystal

Techniques Used: End-sequence Profiling

16) Product Images from "Design, Synthesis, and Evaluation of Alkyl-Quinoxalin-2(1H)-One Derivatives as Anti-Quorum Sensing Molecules, Inhibiting Biofilm Formation in Aeromonas caviae Sch3"

Article Title: Design, Synthesis, and Evaluation of Alkyl-Quinoxalin-2(1H)-One Derivatives as Anti-Quorum Sensing Molecules, Inhibiting Biofilm Formation in Aeromonas caviae Sch3

Journal: Molecules

doi: 10.3390/molecules23123075

Biofilm formation influenced by compounds 9 , 10 , 11 , 12, and 14 in Aeromonas caviae Sch3 at 6 × 10 8 colony forming units (CFU)/mL bacteria. The data are presented as percentages of the biofilm formation of the control, which received no inhibitor treatment and was set as 100%. The compounds were evaluated at 100, 10, and 1 µM concentrations. Dimethyl sulfoxide (DMSO) had no significant effect on biofilm formation; the control experiment data can be found in the Supplementary Materials . Significance was confirmed by the Student’s t -test with an accuracy of * p
Figure Legend Snippet: Biofilm formation influenced by compounds 9 , 10 , 11 , 12, and 14 in Aeromonas caviae Sch3 at 6 × 10 8 colony forming units (CFU)/mL bacteria. The data are presented as percentages of the biofilm formation of the control, which received no inhibitor treatment and was set as 100%. The compounds were evaluated at 100, 10, and 1 µM concentrations. Dimethyl sulfoxide (DMSO) had no significant effect on biofilm formation; the control experiment data can be found in the Supplementary Materials . Significance was confirmed by the Student’s t -test with an accuracy of * p

Techniques Used:

17) Product Images from "An AraC-Type Transcriptional Regulator Encoded on the Enterococcus faecalis Pathogenicity Island Contributes to Pathogenesis and Intracellular Macrophage Survival ▿"

Article Title: An AraC-Type Transcriptional Regulator Encoded on the Enterococcus faecalis Pathogenicity Island Contributes to Pathogenesis and Intracellular Macrophage Survival ▿

Journal: Infection and Immunity

doi: 10.1128/IAI.00930-08

Biofilm density after growth in THB plus 1% glucose. Biofilm formation by E. faecalis strains E99, DBS01, and DBS01(pGT101) on polystyrene microtiter plates was assessed by crystal violet staining. Biofilm assays were performed in triplicate, with 12 replicates for each strain per assay. The error bars represent the means ± the standard errors. A Student t test was used to determine significance levels.
Figure Legend Snippet: Biofilm density after growth in THB plus 1% glucose. Biofilm formation by E. faecalis strains E99, DBS01, and DBS01(pGT101) on polystyrene microtiter plates was assessed by crystal violet staining. Biofilm assays were performed in triplicate, with 12 replicates for each strain per assay. The error bars represent the means ± the standard errors. A Student t test was used to determine significance levels.

Techniques Used: Staining

18) Product Images from "Unsaturated Fatty Acid, cis-2-Decenoic Acid, in Combination with Disinfectants or Antibiotics Removes Pre-Established Biofilms Formed by Food-Related Bacteria"

Article Title: Unsaturated Fatty Acid, cis-2-Decenoic Acid, in Combination with Disinfectants or Antibiotics Removes Pre-Established Biofilms Formed by Food-Related Bacteria

Journal: PLoS ONE

doi: 10.1371/journal.pone.0101677

Effect of CDA combined antimicrobial treatments on killing of pre-established biofilms. CDA treatment reverses biofilm formation in pre-established biofilms and cells remaining on the surface are easily removed and killed various disinfectants (Epimax S and Percidine) or antibiotics (vancomycin; Van, ampicillin; Amp, ciprofloxacin; Cip) in biofilms grown in continuous culture flow cells. Pre-established biofilms were grown for 48 h without any treatment, then were treated with indicated concentrations of antimicrobials alone (- CDA) or combined with 310 nM CDA (+CDA) for 1 h and stained with LIVE/DEAD staining to allow analysis using fluorescence microscopy. The images show microscopic pictures of the biofilms on the surface of cover slip after combinatorial treatments. Images are top-down views (x-y plane); scale bars: 50 µm. Results are representative of 3 separate experiments.
Figure Legend Snippet: Effect of CDA combined antimicrobial treatments on killing of pre-established biofilms. CDA treatment reverses biofilm formation in pre-established biofilms and cells remaining on the surface are easily removed and killed various disinfectants (Epimax S and Percidine) or antibiotics (vancomycin; Van, ampicillin; Amp, ciprofloxacin; Cip) in biofilms grown in continuous culture flow cells. Pre-established biofilms were grown for 48 h without any treatment, then were treated with indicated concentrations of antimicrobials alone (- CDA) or combined with 310 nM CDA (+CDA) for 1 h and stained with LIVE/DEAD staining to allow analysis using fluorescence microscopy. The images show microscopic pictures of the biofilms on the surface of cover slip after combinatorial treatments. Images are top-down views (x-y plane); scale bars: 50 µm. Results are representative of 3 separate experiments.

Techniques Used: Flow Cytometry, Staining, Fluorescence, Microscopy

Induction of planktonic mode of growth in pre-established biofilms formed by food pathogens using CDA. (A) Biofilms were grown for 5 days in petri dishes in which the medium was replaced every 24 h. Dispersion induction was tested by replacing the growth medium with fresh medium containing three different concentrations of CDA (100, 310 or 620 nM) or just the carrier as a control and the cells were incubated for a further 1 h. Medium containing dispersed cells was then homogenized and cell density was determined by measuring the optical density. (B) After 5 days of biofilm growth in flow cell continuous cultures, the influent medium was switched from fresh medium in the test lines to three indicated concentrations of CDA and control lines were switched to new lines containing just the carrier. Effluent runoffs were then collected and cell density was determined by measuring the OD. Error bars indicate standard errors (n = 3) and mean values sharing at least one common lowercase letter shown above the bars are not significantly different ( P-value
Figure Legend Snippet: Induction of planktonic mode of growth in pre-established biofilms formed by food pathogens using CDA. (A) Biofilms were grown for 5 days in petri dishes in which the medium was replaced every 24 h. Dispersion induction was tested by replacing the growth medium with fresh medium containing three different concentrations of CDA (100, 310 or 620 nM) or just the carrier as a control and the cells were incubated for a further 1 h. Medium containing dispersed cells was then homogenized and cell density was determined by measuring the optical density. (B) After 5 days of biofilm growth in flow cell continuous cultures, the influent medium was switched from fresh medium in the test lines to three indicated concentrations of CDA and control lines were switched to new lines containing just the carrier. Effluent runoffs were then collected and cell density was determined by measuring the OD. Error bars indicate standard errors (n = 3) and mean values sharing at least one common lowercase letter shown above the bars are not significantly different ( P-value

Techniques Used: Incubation, Flow Cytometry

Effect of CDA combined antimicrobial treatments on eradication and killing of pre-established biofilms. (A) After 120 h of growth on the surface of SS discs, biofilms were treated for 1 h with biocides alone or combined with 310 nM CDA; CFU plate counts were then performed to assess the viability of the bacteria. (B) The amount of biofilm remaining was determined by the absorbance at 590 nm of crystal violet after staining the 120 h different biofilms in a microtiter plate assay after treatment with tested concentrations of antibiotics alone (- CDA) or in combination with 310 nM CDA (+CDA) for 1 h. All readings are corrected to reflect 0% and 100% controls (blank well, 0%; biofilms without any treatments, 100%). Error bars indicate standard errors (n = 3) and mean values sharing at least one common lowercase letter shown above the bars are not significantly different ( P-value
Figure Legend Snippet: Effect of CDA combined antimicrobial treatments on eradication and killing of pre-established biofilms. (A) After 120 h of growth on the surface of SS discs, biofilms were treated for 1 h with biocides alone or combined with 310 nM CDA; CFU plate counts were then performed to assess the viability of the bacteria. (B) The amount of biofilm remaining was determined by the absorbance at 590 nm of crystal violet after staining the 120 h different biofilms in a microtiter plate assay after treatment with tested concentrations of antibiotics alone (- CDA) or in combination with 310 nM CDA (+CDA) for 1 h. All readings are corrected to reflect 0% and 100% controls (blank well, 0%; biofilms without any treatments, 100%). Error bars indicate standard errors (n = 3) and mean values sharing at least one common lowercase letter shown above the bars are not significantly different ( P-value

Techniques Used: Staining

Effect of CDA combined disinfectant or antibiotic treatments on biofilms surface area. Following dispersion of biofilms by CDA, cells remaining on the surface are easily killed and removed by various disinfectants (Epimax S and Percidine) or antibiotics (vancomycin; Van, ampicillin; Amp, ciprofloxacin; Cip) in biofilms grown in continuous culture flow cells. Pre-established biofilms were grown for 48 h without any treatment and then were treated with indicated concentrations of antimicrobials alone (- CDA) or combined with 310 nM CDA (+ CDA) for 1 h, stained with LIVE/DEAD staining and quantified (percent surface coverage) using digital image analysis. The bars show the levels of biofilm biomass after treatment with antimicrobials alone or combined with 310 nM CDA. Error bars indicate standard errors (n = 3) and mean values sharing at least one common lowercase letter shown above the bars are not significantly different ( P-value
Figure Legend Snippet: Effect of CDA combined disinfectant or antibiotic treatments on biofilms surface area. Following dispersion of biofilms by CDA, cells remaining on the surface are easily killed and removed by various disinfectants (Epimax S and Percidine) or antibiotics (vancomycin; Van, ampicillin; Amp, ciprofloxacin; Cip) in biofilms grown in continuous culture flow cells. Pre-established biofilms were grown for 48 h without any treatment and then were treated with indicated concentrations of antimicrobials alone (- CDA) or combined with 310 nM CDA (+ CDA) for 1 h, stained with LIVE/DEAD staining and quantified (percent surface coverage) using digital image analysis. The bars show the levels of biofilm biomass after treatment with antimicrobials alone or combined with 310 nM CDA. Error bars indicate standard errors (n = 3) and mean values sharing at least one common lowercase letter shown above the bars are not significantly different ( P-value

Techniques Used: Flow Cytometry, Staining

19) Product Images from "Assessment of in vivo versus in vitro biofilm formation of clinical methicillin-resistant Staphylococcus aureus isolates from endotracheal tubes"

Article Title: Assessment of in vivo versus in vitro biofilm formation of clinical methicillin-resistant Staphylococcus aureus isolates from endotracheal tubes

Journal: Scientific Reports

doi: 10.1038/s41598-018-30494-7

Biofilm production dynamics after extubation in 9 ETT-MRSA isolates under O 2 or 5% CO 2 Each color-bar represents the biofilm production of each ETT-MRSA isolate compared with the MRSA-in over days 1–8 after extubation. ( A ) Biofilm production dynamics under O 2 . Maximum biofilm production was on day 2. ( B ) Biofilm production dynamics under CO 2 . Maximum biofilm production was on day 1. Since the 5% CO 2 atmosphere better mimics the atmospheric conditions of mechanical ventilation. When ETT-MRSA are rapidly switched from the ETT environment to O 2 alone, they would need a day to adapt their metabolism to the new atmospheric conditions. Black points represent median biofilm production of the 9 isolates each day. Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Biofilm production dynamics after extubation in 9 ETT-MRSA isolates under O 2 or 5% CO 2 Each color-bar represents the biofilm production of each ETT-MRSA isolate compared with the MRSA-in over days 1–8 after extubation. ( A ) Biofilm production dynamics under O 2 . Maximum biofilm production was on day 2. ( B ) Biofilm production dynamics under CO 2 . Maximum biofilm production was on day 1. Since the 5% CO 2 atmosphere better mimics the atmospheric conditions of mechanical ventilation. When ETT-MRSA are rapidly switched from the ETT environment to O 2 alone, they would need a day to adapt their metabolism to the new atmospheric conditions. Black points represent median biofilm production of the 9 isolates each day. Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used:

Representative scanning electron microscopy of in vivo MRSA biofilm ( A ) Isolate 1 showing an in vivo detached biofilm at low magnification. Sometimes the sample processing for scanning electron microscopy released the biofilm cluster from the endotracheal tube surface. At higher magnification ( B ), cocci morphologies can be distinguished. The pig from which we obtained Isolate 1 was treated with vancomycin. ( C ) Isolate 45 (from a placebo treated pig) showing an in vivo biofilm attached to the endotracheal tube at low magnification. ( D ) at higher magnification, a cocci biofilm cluster was found (white arrow). Abbreviations: MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Representative scanning electron microscopy of in vivo MRSA biofilm ( A ) Isolate 1 showing an in vivo detached biofilm at low magnification. Sometimes the sample processing for scanning electron microscopy released the biofilm cluster from the endotracheal tube surface. At higher magnification ( B ), cocci morphologies can be distinguished. The pig from which we obtained Isolate 1 was treated with vancomycin. ( C ) Isolate 45 (from a placebo treated pig) showing an in vivo biofilm attached to the endotracheal tube at low magnification. ( D ) at higher magnification, a cocci biofilm cluster was found (white arrow). Abbreviations: MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used: Electron Microscopy, In Vivo

Biofilm production of 52 ETT-MRSA isolates compared with the MRSA-in under ambient air or ambient air with 5% CO 2. Each bar represents the biofilm production of each ETT-MRSA isolate versus the MRSA-in (black bar). ( A ) Biofilm production O 2 on day 2 after extubation (peak production); 50% of ETT-MRSA isolates increased biofilm production more than double that of MRSA-in. ( B ) Biofilm production under 5% CO 2 on day 1 after extubation (peak production); 40% of ETT-MRSA isolates increased more than twice MRSA-in biofilm production. The highest biofilm producers ETT-MRSA isolates (n = 9), in dark gray, under both O 2 and 5% CO 2 were selected to undergo the biofilm production dynamics post-extubation. Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Biofilm production of 52 ETT-MRSA isolates compared with the MRSA-in under ambient air or ambient air with 5% CO 2. Each bar represents the biofilm production of each ETT-MRSA isolate versus the MRSA-in (black bar). ( A ) Biofilm production O 2 on day 2 after extubation (peak production); 50% of ETT-MRSA isolates increased biofilm production more than double that of MRSA-in. ( B ) Biofilm production under 5% CO 2 on day 1 after extubation (peak production); 40% of ETT-MRSA isolates increased more than twice MRSA-in biofilm production. The highest biofilm producers ETT-MRSA isolates (n = 9), in dark gray, under both O 2 and 5% CO 2 were selected to undergo the biofilm production dynamics post-extubation. Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used:

Representative confocal laser scanning microscopy of in vivo MRSA biofilm Biofilm clusters (white arrows) were stained with the LIVE/DEAD BacLight kit (INVITROGEN, Barcelona, Spain). Viable bacteria (stained green by SYTO 9) are visible, but dead bacteria (stained red by propidium iodide) were infrequently detected. The nuclei and cytoplasm of eukaryotic cells from the pig were also stained nonspecifically by propidium iodide and SYTO 9 (large red and green blotches, respectively). ( A , B ) correspond to the in vivo biofilms of Isolate 39 and Isolate 45 obtained from pigs treated with vancomycin and placebo, respectively. Abbreviations: MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Representative confocal laser scanning microscopy of in vivo MRSA biofilm Biofilm clusters (white arrows) were stained with the LIVE/DEAD BacLight kit (INVITROGEN, Barcelona, Spain). Viable bacteria (stained green by SYTO 9) are visible, but dead bacteria (stained red by propidium iodide) were infrequently detected. The nuclei and cytoplasm of eukaryotic cells from the pig were also stained nonspecifically by propidium iodide and SYTO 9 (large red and green blotches, respectively). ( A , B ) correspond to the in vivo biofilms of Isolate 39 and Isolate 45 obtained from pigs treated with vancomycin and placebo, respectively. Abbreviations: MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used: Confocal Laser Scanning Microscopy, In Vivo, Staining

Effect of systemic antibiotic treatment on biofilm production in the 52 ETT-MRSA isolates under O 2 or 5% CO 2 conditions Median (interquartile range) values for biofilm production of the 52 ETT-MRSA compared with the MRSA-in under O 2 ( A ) and 5% CO 2 ( B ). Time of assessment: day of peak production. Biofilm production was not influenced by systemic treatment with placebo (n = 19), linezolid (n = 11), or vancomycin (n = 22) under either O 2 (1.96 [0.61–3.02], 2.30 [0.64–3.83], and 1.49 [0.63–2.67], respectively; p = 0.92) or 5% CO 2 (2.02 [1.16–2.34], 1.36 [0.48–2.75], and 1.09 [0.34–2.38], respectively; p = 0.62). Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .
Figure Legend Snippet: Effect of systemic antibiotic treatment on biofilm production in the 52 ETT-MRSA isolates under O 2 or 5% CO 2 conditions Median (interquartile range) values for biofilm production of the 52 ETT-MRSA compared with the MRSA-in under O 2 ( A ) and 5% CO 2 ( B ). Time of assessment: day of peak production. Biofilm production was not influenced by systemic treatment with placebo (n = 19), linezolid (n = 11), or vancomycin (n = 22) under either O 2 (1.96 [0.61–3.02], 2.30 [0.64–3.83], and 1.49 [0.63–2.67], respectively; p = 0.92) or 5% CO 2 (2.02 [1.16–2.34], 1.36 [0.48–2.75], and 1.09 [0.34–2.38], respectively; p = 0.62). Abbreviations: 5% CO 2 , ambient air with 5% CO 2 ; O 2 , ambient air; ETT-MRSA, clinical MRSA isolates from endotracheal tubes; MRSA-in, MRSA inoculated into pigs’ lungs; MRSA, methicillin-resistant Staphylococcus aureus .

Techniques Used:

20) Product Images from "Fatty Acid Potassium Had Beneficial Bactericidal Effects and Removed Staphylococcus aureus Biofilms while Exhibiting Reduced Cytotoxicity towards Mouse Fibroblasts and Human Keratinocytes"

Article Title: Fatty Acid Potassium Had Beneficial Bactericidal Effects and Removed Staphylococcus aureus Biofilms while Exhibiting Reduced Cytotoxicity towards Mouse Fibroblasts and Human Keratinocytes

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms20020312

Percentage MRSA OJ-1 biofilm removal values of various types of fatty acid potassium, SLES, and SLS. The formed biofilm was placed in contact with the test substance for 1 min. The biofilm remaining in the well was stained with 0.1% crystal violet and quantified based on the absorbance at 570 nm using a microplate reader. The results are expressed as mean ± SD values ( n = 6). * p
Figure Legend Snippet: Percentage MRSA OJ-1 biofilm removal values of various types of fatty acid potassium, SLES, and SLS. The formed biofilm was placed in contact with the test substance for 1 min. The biofilm remaining in the well was stained with 0.1% crystal violet and quantified based on the absorbance at 570 nm using a microplate reader. The results are expressed as mean ± SD values ( n = 6). * p

Techniques Used: Staining

Scanning electron microscopy. Scanning electron microscopy micrographs of the untreated ( a ) and C18:1K-treated ( b ) biofilms that formed on the test pieces (2000×). MRSA biofilms formed on the test pieces after they had been incubated for 24 h at 37 °C. The MRSA biofilms that formed on the test pieces were removed by C18:1K. ( a ) An untreated biofilm that formed on a test piece; ( b ) A biofilm that had been treated with C18:1K for 1 min.
Figure Legend Snippet: Scanning electron microscopy. Scanning electron microscopy micrographs of the untreated ( a ) and C18:1K-treated ( b ) biofilms that formed on the test pieces (2000×). MRSA biofilms formed on the test pieces after they had been incubated for 24 h at 37 °C. The MRSA biofilms that formed on the test pieces were removed by C18:1K. ( a ) An untreated biofilm that formed on a test piece; ( b ) A biofilm that had been treated with C18:1K for 1 min.

Techniques Used: Electron Microscopy, Incubation

21) Product Images from "Effects of Signal Disruption Depends on the Substrate Preference of the Lactonase"

Article Title: Effects of Signal Disruption Depends on the Substrate Preference of the Lactonase

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2019.03003

Lactonase treatment is not cytotoxic, acts in a dose dependent manner, and is effective at inhibiting biofilm formation in Pseudomonas aeruginosa . (A) Cell density of P. aeruginosa PA14, without treatment (control; dark gray bar), or with addition of 5-FU (5′-fluorouracil; 60 μM; light gray bar), SsoPox mutant 5A8 (an inactive lactonase; 125 μg/mL; black bar), Ssopox W263I (a lactonase; 125 μg/mL; red bar), or GcL (a lactonase; 55 μg/mL; blue bar). Statistical analysis was performed using Student’s t test. (B) Normalized PA14 biofilm quantity as quantified by Crystal Violet assay, using varying concentrations of Ssopox W263I (red bars), GcL (blue bars), or controls 5-FU (fluorouracil; 60 μM; gray bar), or BSA (bovine serum albumin; 100 μg/mL; black bar). Concentrations of Ssopox W263I and GcL are μg/mL. Statistical analysis was performed using Student’s t test. ∗ indicate statistical significance. Maximal inhibition of biofilm was at 125 μg/mL for Ssopox, and 55 μg/mL for GcL.
Figure Legend Snippet: Lactonase treatment is not cytotoxic, acts in a dose dependent manner, and is effective at inhibiting biofilm formation in Pseudomonas aeruginosa . (A) Cell density of P. aeruginosa PA14, without treatment (control; dark gray bar), or with addition of 5-FU (5′-fluorouracil; 60 μM; light gray bar), SsoPox mutant 5A8 (an inactive lactonase; 125 μg/mL; black bar), Ssopox W263I (a lactonase; 125 μg/mL; red bar), or GcL (a lactonase; 55 μg/mL; blue bar). Statistical analysis was performed using Student’s t test. (B) Normalized PA14 biofilm quantity as quantified by Crystal Violet assay, using varying concentrations of Ssopox W263I (red bars), GcL (blue bars), or controls 5-FU (fluorouracil; 60 μM; gray bar), or BSA (bovine serum albumin; 100 μg/mL; black bar). Concentrations of Ssopox W263I and GcL are μg/mL. Statistical analysis was performed using Student’s t test. ∗ indicate statistical significance. Maximal inhibition of biofilm was at 125 μg/mL for Ssopox, and 55 μg/mL for GcL.

Techniques Used: Mutagenesis, Crystal Violet Assay, Inhibition

Data bar table showing effectiveness of lactonase treatment on measured virulence factors (VFs). Data bar table showing the effects of Ssopox (red background) or GcL (blue background) on four virulence factors produced by P. aeruginosa clinical isolates (39), including biofilm, elastase, protease, pyocyanin. Data bars show inhibition (rightward deflection) or stimulation (leftward deflection). Statistically significant deflections ( p
Figure Legend Snippet: Data bar table showing effectiveness of lactonase treatment on measured virulence factors (VFs). Data bar table showing the effects of Ssopox (red background) or GcL (blue background) on four virulence factors produced by P. aeruginosa clinical isolates (39), including biofilm, elastase, protease, pyocyanin. Data bars show inhibition (rightward deflection) or stimulation (leftward deflection). Statistically significant deflections ( p

Techniques Used: Produced, Inhibition

22) Product Images from "Acinetobacter baumannii biofilm biomass mediates tolerance to cold plasma"

Article Title: Acinetobacter baumannii biofilm biomass mediates tolerance to cold plasma

Journal: Letters in Applied Microbiology

doi: 10.1111/lam.13122

Survival curves for Acinetobacter b aumannii biofilms grown for (a) 24 h, (b) 48 h and (c) 72 h respectively. (d) Exhibits log reduction of viable for biofilms grown for 24 h (black bars), 48 h (white bars) and 72 h (grey bars). Each bar represents the mean of three replicates with the standard error of the mean. (e) Demonstrates biomass of A. baumannii biofilms grown for 24 h (black bars), 48 h (white bars) and 72 h (grey bars). Each bar represents the mean of 12 replicates with error bars representing the standard error of the mean. One‐way anova with Tukey's multiple comparisons test performed. **** P value
Figure Legend Snippet: Survival curves for Acinetobacter b aumannii biofilms grown for (a) 24 h, (b) 48 h and (c) 72 h respectively. (d) Exhibits log reduction of viable for biofilms grown for 24 h (black bars), 48 h (white bars) and 72 h (grey bars). Each bar represents the mean of three replicates with the standard error of the mean. (e) Demonstrates biomass of A. baumannii biofilms grown for 24 h (black bars), 48 h (white bars) and 72 h (grey bars). Each bar represents the mean of 12 replicates with error bars representing the standard error of the mean. One‐way anova with Tukey's multiple comparisons test performed. **** P value

Techniques Used:

Scanning electron microscopy images ( SEM ) of an unexposed 24‐h Acinetobacter b aumannii biofilm (a) and (c) and a 6‐min, plasma‐treated A. baumannii biofilm (b) and (d) captured using a JEOL field emission SEM JSM ‐6500F.
Figure Legend Snippet: Scanning electron microscopy images ( SEM ) of an unexposed 24‐h Acinetobacter b aumannii biofilm (a) and (c) and a 6‐min, plasma‐treated A. baumannii biofilm (b) and (d) captured using a JEOL field emission SEM JSM ‐6500F.

Techniques Used: Electron Microscopy

23) Product Images from "Design, Synthesis, and Evaluation of Alkyl-Quinoxalin-2(1H)-One Derivatives as Anti-Quorum Sensing Molecules, Inhibiting Biofilm Formation in Aeromonas caviae Sch3"

Article Title: Design, Synthesis, and Evaluation of Alkyl-Quinoxalin-2(1H)-One Derivatives as Anti-Quorum Sensing Molecules, Inhibiting Biofilm Formation in Aeromonas caviae Sch3

Journal: Molecules

doi: 10.3390/molecules23123075

2.3. Inhibition of Biofilm Formation by Anti-Quorum Sensing Compounds
Figure Legend Snippet: 2.3. Inhibition of Biofilm Formation by Anti-Quorum Sensing Compounds

Techniques Used: Inhibition

24) Product Images from "Antibacterial and Antibiofilm Activities of Nonpolar Extracts of Allium stipitatum Regel. against Multidrug Resistant Bacteria"

Article Title: Antibacterial and Antibiofilm Activities of Nonpolar Extracts of Allium stipitatum Regel. against Multidrug Resistant Bacteria

Journal: BioMed Research International

doi: 10.1155/2018/9845075

Representative confocal images of mature biofilms and ASHE and ASDE treated biofilms. Panels ( c–h ) represent ASHE and ASDE treated groups of MSSA. Panels ( k–p ) represent ASHE and ASDE treated groups of MRSA. Panels ( s–x ) represent ASHE and ASDE treated groups of A. baumannii . Panels ( aa–af) represent ASHE and ASDE treated groups of S. maltophilia . Micrographs of fluorescent biofilms were obtained by confocal microscopy.
Figure Legend Snippet: Representative confocal images of mature biofilms and ASHE and ASDE treated biofilms. Panels ( c–h ) represent ASHE and ASDE treated groups of MSSA. Panels ( k–p ) represent ASHE and ASDE treated groups of MRSA. Panels ( s–x ) represent ASHE and ASDE treated groups of A. baumannii . Panels ( aa–af) represent ASHE and ASDE treated groups of S. maltophilia . Micrographs of fluorescent biofilms were obtained by confocal microscopy.

Techniques Used: Confocal Microscopy

Effect of ASHE and ASDE on the viability of ( a ) S. aureus , ( b ) MRSA, ( c ) A. baumannii , and ( d ) S. maltophilia biofilms at concentrations of 1x, 2x, and 4x MICs with control (0x MIC). Comparison of absorbance between control and treated samples at 490 nm by XTT assay. ∗∗∗ p
Figure Legend Snippet: Effect of ASHE and ASDE on the viability of ( a ) S. aureus , ( b ) MRSA, ( c ) A. baumannii , and ( d ) S. maltophilia biofilms at concentrations of 1x, 2x, and 4x MICs with control (0x MIC). Comparison of absorbance between control and treated samples at 490 nm by XTT assay. ∗∗∗ p

Techniques Used: XTT Assay

25) Product Images from "Pulmonary Surfactant Promotes Virulence Gene Expression and Biofilm Formation in Klebsiella pneumoniae"

Article Title: Pulmonary Surfactant Promotes Virulence Gene Expression and Biofilm Formation in Klebsiella pneumoniae

Journal: Infection and Immunity

doi: 10.1128/IAI.00135-18

qRT-PCR validation of induced transcripts in K. pneumoniae following exposure to lung surfactant. The relative abundance of seven Survanta-induced transcripts detected through the microarray were reexamined using quantitative RT-PCR with RNA collected from three independent Survanta induction experiments as described in the Materials and Methods section. Genes for analysis were chosen to represent a range of cellular functions, including lipid metabolism ( fadB and Kpn_02053 [ dhcA ]), biofilm formation ( mrkA ), branched-chain amino acid synthesis ( leuA ), nutrient uptake ( proV ), polyamine efflux ( mdtJ ), and LPS modification ( arnA ). Raw transcript expression values were normalized to those for Kpn_04184 , which exhibited no change in expression between conditions in our microarrays. We examined expression in both MGH 78578 (M) and KPPR1 (K), though our primers designed for fadB and Kpn_02053 in MGH 78578 were not usable in KPPR1 due to multiple products (noted with the # symbol). Statistical analysis was conducted via two-way analysis of variance with a Sidak posttest analyzing in-strain changes comparing expression under conditions with Survanta additions to expression under the lactate-alone condition. The data shown summarize those from three independent Survanta induction experiments, and statistical significance is indicated as follows: *, P
Figure Legend Snippet: qRT-PCR validation of induced transcripts in K. pneumoniae following exposure to lung surfactant. The relative abundance of seven Survanta-induced transcripts detected through the microarray were reexamined using quantitative RT-PCR with RNA collected from three independent Survanta induction experiments as described in the Materials and Methods section. Genes for analysis were chosen to represent a range of cellular functions, including lipid metabolism ( fadB and Kpn_02053 [ dhcA ]), biofilm formation ( mrkA ), branched-chain amino acid synthesis ( leuA ), nutrient uptake ( proV ), polyamine efflux ( mdtJ ), and LPS modification ( arnA ). Raw transcript expression values were normalized to those for Kpn_04184 , which exhibited no change in expression between conditions in our microarrays. We examined expression in both MGH 78578 (M) and KPPR1 (K), though our primers designed for fadB and Kpn_02053 in MGH 78578 were not usable in KPPR1 due to multiple products (noted with the # symbol). Statistical analysis was conducted via two-way analysis of variance with a Sidak posttest analyzing in-strain changes comparing expression under conditions with Survanta additions to expression under the lactate-alone condition. The data shown summarize those from three independent Survanta induction experiments, and statistical significance is indicated as follows: *, P

Techniques Used: Quantitative RT-PCR, Microarray, Modification, Expressing

26) Product Images from "The Ability of Lytic Staphylococcal Podovirus vB_SauP_phiAGO1.3 to Coexist in Equilibrium With Its Host Facilitates the Selection of Host Mutants of Attenuated Virulence but Does Not Preclude the Phage Antistaphylococcal Activity in a Nematode Infection Model"

Article Title: The Ability of Lytic Staphylococcal Podovirus vB_SauP_phiAGO1.3 to Coexist in Equilibrium With Its Host Facilitates the Selection of Host Mutants of Attenuated Virulence but Does Not Preclude the Phage Antistaphylococcal Activity in a Nematode Infection Model

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.03227

Activity of bacteriophage phiAGO1.3 in lysis of Staphylococcus aureus strains of various clonal complexes and in the destruction of their biofilms. Strain numbers are as in the MICROBANK collection, with the exception of laboratory strain RN4220. Environmental and bovine mastitis S. aureus isolates are marked by (E) and (M), respectively. Density changes of liquid cultures of S. aureus upon addition of phiAGO1.3 are exemplified in Supplementary Figure 3 .
Figure Legend Snippet: Activity of bacteriophage phiAGO1.3 in lysis of Staphylococcus aureus strains of various clonal complexes and in the destruction of their biofilms. Strain numbers are as in the MICROBANK collection, with the exception of laboratory strain RN4220. Environmental and bovine mastitis S. aureus isolates are marked by (E) and (M), respectively. Density changes of liquid cultures of S. aureus upon addition of phiAGO1.3 are exemplified in Supplementary Figure 3 .

Techniques Used: Activity Assay, Lysis

27) Product Images from "Synergistic Interactions within a Multispecies Biofilm Enhance Individual Species Protection against Grazing by a Pelagic Protozoan"

Article Title: Synergistic Interactions within a Multispecies Biofilm Enhance Individual Species Protection against Grazing by a Pelagic Protozoan

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2017.02649

Change in viable cell numbers from the biofilm fractions of mono and multispecies cultures grazed with T. pyriformis after 24 and 96 h obtained from plating. The data points indicate the change in cell numbers of grazed biofilm fraction relative to the non-grazed biofilm fraction ± SEM obtained from two biological replicates.
Figure Legend Snippet: Change in viable cell numbers from the biofilm fractions of mono and multispecies cultures grazed with T. pyriformis after 24 and 96 h obtained from plating. The data points indicate the change in cell numbers of grazed biofilm fraction relative to the non-grazed biofilm fraction ± SEM obtained from two biological replicates.

Techniques Used:

Impact of grazing by T. pyriformis on the population dynamics of the individual bacterial species in the multispecies consortia as assessed by qPCR. Cell numbers of the individual bacterial members in (A) the multispecies biofilm fraction and (B) the multispecies planktonic fraction after 24 h of grazing.
Figure Legend Snippet: Impact of grazing by T. pyriformis on the population dynamics of the individual bacterial species in the multispecies consortia as assessed by qPCR. Cell numbers of the individual bacterial members in (A) the multispecies biofilm fraction and (B) the multispecies planktonic fraction after 24 h of grazing.

Techniques Used: Real-time Polymerase Chain Reaction

Grazing for 24 h by protozoa on the four-species mixed cultures. FISH based staining and confocal imaging shows the distribution of the different bacterial species in and around the protozoan cells. Applying the fluorescence filter channels, it is observed that X. retroflexus is abundantly present within the food vacuoles (indicated by the arrows) of T. pyriformis (A) . S. rhizophila (B) is detected to a lesser extent whereas M. oxydans (C) and P. amylolyticus (D) cells are not visibly present in the food vacuoles. (E,F) Depict the overlay and bright-field images, respectively. Panels (G,H) were included to phase out the dominating fluorescence signals from X. retroflexus and visualize the other bacterial members in the biofilm consortia around the ciliate.
Figure Legend Snippet: Grazing for 24 h by protozoa on the four-species mixed cultures. FISH based staining and confocal imaging shows the distribution of the different bacterial species in and around the protozoan cells. Applying the fluorescence filter channels, it is observed that X. retroflexus is abundantly present within the food vacuoles (indicated by the arrows) of T. pyriformis (A) . S. rhizophila (B) is detected to a lesser extent whereas M. oxydans (C) and P. amylolyticus (D) cells are not visibly present in the food vacuoles. (E,F) Depict the overlay and bright-field images, respectively. Panels (G,H) were included to phase out the dominating fluorescence signals from X. retroflexus and visualize the other bacterial members in the biofilm consortia around the ciliate.

Techniques Used: Fluorescence In Situ Hybridization, Staining, Imaging, Fluorescence

Biofilm forming index (BFI) of mono and mixed species cultures subject to Tetrahymena pyriformis (Tp) grazing and non-grazed cultures at 12, 24, and 96 h. The data points indicate the biofilm mean ± standard error of the mean (SEM) obtained from five biological replicates. ∗ P
Figure Legend Snippet: Biofilm forming index (BFI) of mono and mixed species cultures subject to Tetrahymena pyriformis (Tp) grazing and non-grazed cultures at 12, 24, and 96 h. The data points indicate the biofilm mean ± standard error of the mean (SEM) obtained from five biological replicates. ∗ P

Techniques Used:

Biofilm formation in the presence of T. pyriformis in three-species bacterial consortia. X. retroflexus is vital for biofilm development. Biofilm fold was calculated as the ratio of Abs 590 [(three – species biofilm cultured with grazer cells + SEM) – (three – species biofilm as control – SEM)] to Abs 590 (three – species biofilm cultured with grazer cells + SEM).
Figure Legend Snippet: Biofilm formation in the presence of T. pyriformis in three-species bacterial consortia. X. retroflexus is vital for biofilm development. Biofilm fold was calculated as the ratio of Abs 590 [(three – species biofilm cultured with grazer cells + SEM) – (three – species biofilm as control – SEM)] to Abs 590 (three – species biofilm cultured with grazer cells + SEM).

Techniques Used: Cell Culture

28) Product Images from "A Recombinant Snake Cathelicidin Derivative Peptide: Antibiofilm Properties and Expression in Escherichia coli"

Article Title: A Recombinant Snake Cathelicidin Derivative Peptide: Antibiofilm Properties and Expression in Escherichia coli

Journal: Biomolecules

doi: 10.3390/biom8040118

The activities of levofloxacin and Cath-A against the established biofilms of P. aeruginisa ( a ) and A . baumannii ( b ) at different concentrations (4–256 µg/mL). The antibiofilm effect of the peptide and levofloxacin was assessed by cv staining after 24 h of incubation. Biofilm formation was determined by measuring the absorbance at 595 nm using a microplate reader. Controls were untreated inoculated bacteria in Mueller Hinton (MH) broth. Data are shown as the mean ± SD of three independent tests. Significant different values were (# p
Figure Legend Snippet: The activities of levofloxacin and Cath-A against the established biofilms of P. aeruginisa ( a ) and A . baumannii ( b ) at different concentrations (4–256 µg/mL). The antibiofilm effect of the peptide and levofloxacin was assessed by cv staining after 24 h of incubation. Biofilm formation was determined by measuring the absorbance at 595 nm using a microplate reader. Controls were untreated inoculated bacteria in Mueller Hinton (MH) broth. Data are shown as the mean ± SD of three independent tests. Significant different values were (# p

Techniques Used: Staining, Incubation

29) Product Images from "RNA-Seq-based transcriptome analysis of methicillin-resistant Staphylococcus aureus biofilm inhibition by ursolic acid and resveratrol"

Article Title: RNA-Seq-based transcriptome analysis of methicillin-resistant Staphylococcus aureus biofilm inhibition by ursolic acid and resveratrol

Journal: Scientific Reports

doi: 10.1038/srep05467

Distribution of differentially expressed genes in inhibitory and removed conditions Red, yellow, and blue correspond to genes with high, medium, and low expression, respectively. (a) Distribution of differentially expressed genes in the resveratrol inhibiting methicillin-resistant Staphylococcus aureus (MRSA) biofilm formation condition. From outside to inside, the three circles in each plot correspond to the following: (1 and 2) log2 of the fragments per kilobase of transcript per million mapped fragments (FPKM) of each gene in MR100 and M18C06; (3) genes differentially expressed in MR100 compared with M18C06. (b) Distribution of the differentially expressed genes in ursolic acid inhibitory MRSA biofilm formation condition. From outside to inside, the three circles in each plot correspond to the following: (1 and 2) log2 of FPKM each gene in MU30 and M18C06, respectively; (3) genes differentially expressed in MU30 compared with M18C06. (c) Distribution of the differentially expressed genes in the resveratrol and its combination with vancomycin removed established MRSA biofilm conditions. From outside to inside, the six circles in each plot correspond to the following: (1, 2, and 3) log2 of FPKM of each gene in MVR, MR150, and M36C05, respectively; (4) genes differentially expressed in MVR compared with MR150; (5 and 6) genes differentially expressed in MVR and MR150 compared with M36C05, respectively.
Figure Legend Snippet: Distribution of differentially expressed genes in inhibitory and removed conditions Red, yellow, and blue correspond to genes with high, medium, and low expression, respectively. (a) Distribution of differentially expressed genes in the resveratrol inhibiting methicillin-resistant Staphylococcus aureus (MRSA) biofilm formation condition. From outside to inside, the three circles in each plot correspond to the following: (1 and 2) log2 of the fragments per kilobase of transcript per million mapped fragments (FPKM) of each gene in MR100 and M18C06; (3) genes differentially expressed in MR100 compared with M18C06. (b) Distribution of the differentially expressed genes in ursolic acid inhibitory MRSA biofilm formation condition. From outside to inside, the three circles in each plot correspond to the following: (1 and 2) log2 of FPKM each gene in MU30 and M18C06, respectively; (3) genes differentially expressed in MU30 compared with M18C06. (c) Distribution of the differentially expressed genes in the resveratrol and its combination with vancomycin removed established MRSA biofilm conditions. From outside to inside, the six circles in each plot correspond to the following: (1, 2, and 3) log2 of FPKM of each gene in MVR, MR150, and M36C05, respectively; (4) genes differentially expressed in MVR compared with MR150; (5 and 6) genes differentially expressed in MVR and MR150 compared with M36C05, respectively.

Techniques Used: Expressing

Heatmap of differentially expressed genes associated with methicillin-resistant Staphylococcus aureus (MRSA) biofilm formation and virulence. (a) Resveratrol inhibiting MRSA biofilm formation condition, (b) ursolic acid inhibiting MRSA biofilm condition, and (c) resveratrol and its combination with vancomycin removing established MRSA biofilm condition. Chemically, resveratrol had no correlation with ursolic acid, so we performed separate differential expression analyses.
Figure Legend Snippet: Heatmap of differentially expressed genes associated with methicillin-resistant Staphylococcus aureus (MRSA) biofilm formation and virulence. (a) Resveratrol inhibiting MRSA biofilm formation condition, (b) ursolic acid inhibiting MRSA biofilm condition, and (c) resveratrol and its combination with vancomycin removing established MRSA biofilm condition. Chemically, resveratrol had no correlation with ursolic acid, so we performed separate differential expression analyses.

Techniques Used: Expressing

Coverage changes of differentially expressed genes associated with methicillin-resistant Staphylococcus aureus (MRSA) biofilm and virulence in removing established MRSA biofilm conditions. Red, blue, and green lines represent M36C05, MVR, and MR150, respectively. For the x-axis, the position of the genome increases from left to right, and the length of each gene is proportional to the width of the position in the x-axis. The use of “..” indicates discontinuous gene segments.
Figure Legend Snippet: Coverage changes of differentially expressed genes associated with methicillin-resistant Staphylococcus aureus (MRSA) biofilm and virulence in removing established MRSA biofilm conditions. Red, blue, and green lines represent M36C05, MVR, and MR150, respectively. For the x-axis, the position of the genome increases from left to right, and the length of each gene is proportional to the width of the position in the x-axis. The use of “..” indicates discontinuous gene segments.

Techniques Used:

30) Product Images from "Fatty Acid Potassium Had Beneficial Bactericidal Effects and Removed Staphylococcus aureus Biofilms while Exhibiting Reduced Cytotoxicity towards Mouse Fibroblasts and Human Keratinocytes"

Article Title: Fatty Acid Potassium Had Beneficial Bactericidal Effects and Removed Staphylococcus aureus Biofilms while Exhibiting Reduced Cytotoxicity towards Mouse Fibroblasts and Human Keratinocytes

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms20020312

Percentage MRSA OJ-1 biofilm removal values of various types of fatty acid potassium, SLES, and SLS. The formed biofilm was placed in contact with the test substance for 1 min. The biofilm remaining in the well was stained with 0.1% crystal violet and quantified based on the absorbance at 570 nm using a microplate reader. The results are expressed as mean ± SD values ( n = 6). * p
Figure Legend Snippet: Percentage MRSA OJ-1 biofilm removal values of various types of fatty acid potassium, SLES, and SLS. The formed biofilm was placed in contact with the test substance for 1 min. The biofilm remaining in the well was stained with 0.1% crystal violet and quantified based on the absorbance at 570 nm using a microplate reader. The results are expressed as mean ± SD values ( n = 6). * p

Techniques Used: Staining

Scanning electron microscopy. Scanning electron microscopy micrographs of the untreated ( a ) and C18:1K-treated ( b ) biofilms that formed on the test pieces (2000×). MRSA biofilms formed on the test pieces after they had been incubated for 24 h at 37 °C. The MRSA biofilms that formed on the test pieces were removed by C18:1K. ( a ) An untreated biofilm that formed on a test piece; ( b ) A biofilm that had been treated with C18:1K for 1 min.
Figure Legend Snippet: Scanning electron microscopy. Scanning electron microscopy micrographs of the untreated ( a ) and C18:1K-treated ( b ) biofilms that formed on the test pieces (2000×). MRSA biofilms formed on the test pieces after they had been incubated for 24 h at 37 °C. The MRSA biofilms that formed on the test pieces were removed by C18:1K. ( a ) An untreated biofilm that formed on a test piece; ( b ) A biofilm that had been treated with C18:1K for 1 min.

Techniques Used: Electron Microscopy, Incubation

31) Product Images from "TolC Promotes ExPEC Biofilm Formation and Curli Production in Response to Medium Osmolarity"

Article Title: TolC Promotes ExPEC Biofilm Formation and Curli Production in Response to Medium Osmolarity

Journal: BioMed Research International

doi: 10.1155/2014/574274

Effect of medium osmolarity on biofilm formation by the WT and Δ tolC strains as determined by crystal violet biofilm assay. Both strains were cultivated in 1/2 M9 medium supplemented with different concentrations of NaCl (a) or sucrose (b) at 28°C for 5 days. Biofilm formation was quantified, and the results are shown as means ± standard error of the mean. Significant differences between the WT and Δ tolC strains were determined using one-tailed unpaired Student's t -test. ∗ P
Figure Legend Snippet: Effect of medium osmolarity on biofilm formation by the WT and Δ tolC strains as determined by crystal violet biofilm assay. Both strains were cultivated in 1/2 M9 medium supplemented with different concentrations of NaCl (a) or sucrose (b) at 28°C for 5 days. Biofilm formation was quantified, and the results are shown as means ± standard error of the mean. Significant differences between the WT and Δ tolC strains were determined using one-tailed unpaired Student's t -test. ∗ P

Techniques Used: Biofilm Production Assay, One-tailed Test

Crystal violet biofilm assay of ExPEC strains grown in M9 or 1/2 M9 media. Biofilms were formed in 96-well plates at 28°C for 5 days and quantified by measuring the OD at 630 nm (OD 630 ) of dissolved crystal violet. Means and standard error of the mean values of OD 630 values of 24 replicate wells from three independent cultures are shown. The cutoff OD (ODc) was defined as three standard deviations above the mean OD of the negative control. Biofilm production was classified as follows: OD
Figure Legend Snippet: Crystal violet biofilm assay of ExPEC strains grown in M9 or 1/2 M9 media. Biofilms were formed in 96-well plates at 28°C for 5 days and quantified by measuring the OD at 630 nm (OD 630 ) of dissolved crystal violet. Means and standard error of the mean values of OD 630 values of 24 replicate wells from three independent cultures are shown. The cutoff OD (ODc) was defined as three standard deviations above the mean OD of the negative control. Biofilm production was classified as follows: OD

Techniques Used: Biofilm Production Assay, Negative Control

32) Product Images from "Achromobacter Species Isolated from Cystic Fibrosis Patients Reveal Distinctly Different Biofilm Morphotypes"

Article Title: Achromobacter Species Isolated from Cystic Fibrosis Patients Reveal Distinctly Different Biofilm Morphotypes

Journal: Microorganisms

doi: 10.3390/microorganisms4030033

Enzymatic treatment both reduced and increased Achromobacter biofilm formation and caused biofilm dispersal. Enzymatic treatment during biofilm formation ( a – c , prevention) and enzymatic treatment of 24 h old biofilms ( d – f , dispersal) in A. xylosoxidans LMG 1863 T , A. xylosoxidans CF2-b and A. insuavis CF4-b. Biofilms were subjected to treatment with alginate lyase ( a and d ), DNase ( b and e ) and subtilisin ( c and f ). 0: untreated, 1: low concentration of enzyme (50 µg/mL), 2: high concentration of enzyme (200 g/mL) * Statistically significant reduction compared to untreated. ** Statistically significant increase compared to untreated. A p -value
Figure Legend Snippet: Enzymatic treatment both reduced and increased Achromobacter biofilm formation and caused biofilm dispersal. Enzymatic treatment during biofilm formation ( a – c , prevention) and enzymatic treatment of 24 h old biofilms ( d – f , dispersal) in A. xylosoxidans LMG 1863 T , A. xylosoxidans CF2-b and A. insuavis CF4-b. Biofilms were subjected to treatment with alginate lyase ( a and d ), DNase ( b and e ) and subtilisin ( c and f ). 0: untreated, 1: low concentration of enzyme (50 µg/mL), 2: high concentration of enzyme (200 g/mL) * Statistically significant reduction compared to untreated. ** Statistically significant increase compared to untreated. A p -value

Techniques Used: Concentration Assay

Mean production of biofilm ± SD in 23 Achromobacter isolates and P. aeruginosa DSMZ 19880 measured by crystal violet staining (strains are listed in Table 1 ).
Figure Legend Snippet: Mean production of biofilm ± SD in 23 Achromobacter isolates and P. aeruginosa DSMZ 19880 measured by crystal violet staining (strains are listed in Table 1 ).

Techniques Used: Staining

Biofilm structure and matrix composition of A. xylosoxisans LMG 1863 T and clinical isolates A. xylosoxidans CF2-b and A. insuavis CF4-b. Biofilms were grown for 48 h at continuous flow ( A – C ) and at static conditions ( D ) and visualized by confocal microscopy. Living cells were stained with Syto 9 (green), dead cells and extracellular DNA with propidium iodide (red) and polysaccharides were stained with fluorescent brightener (blue). A. xylosoxidans LMG 1863 T ( A ) and A. xylosoxisans CF2-b ( B ) produced surface attached biofilm, whereas A. insuavis CF4-b ( C ) failed to form biofilm under continuous flow, but formed suspended aggregates encased in polysaccharides (blue) when grown at static conditions ( D ).
Figure Legend Snippet: Biofilm structure and matrix composition of A. xylosoxisans LMG 1863 T and clinical isolates A. xylosoxidans CF2-b and A. insuavis CF4-b. Biofilms were grown for 48 h at continuous flow ( A – C ) and at static conditions ( D ) and visualized by confocal microscopy. Living cells were stained with Syto 9 (green), dead cells and extracellular DNA with propidium iodide (red) and polysaccharides were stained with fluorescent brightener (blue). A. xylosoxidans LMG 1863 T ( A ) and A. xylosoxisans CF2-b ( B ) produced surface attached biofilm, whereas A. insuavis CF4-b ( C ) failed to form biofilm under continuous flow, but formed suspended aggregates encased in polysaccharides (blue) when grown at static conditions ( D ).

Techniques Used: Flow Cytometry, Confocal Microscopy, Staining, Produced

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Article Title: A Recombinant Snake Cathelicidin Derivative Peptide: Antibiofilm Properties and Expression in Escherichia coli
Article Snippet: .. The absorbance of the stained biofilm was measured at an optical density (OD) of 595 nm by a microplate reader (BioTek Instruments, PowerWave XS, Winooski, VT USA). .. Recombinant Vector Construct The entire gene sequence, including the rec- Cath-A with the additional enterokinase cleavage site was designed, as seen in .

Article Title: Type III Secretion System Translocon Component EseB Forms Filaments on and Mediates Autoaggregation of and Biofilm Formation by Edwardsiella tarda
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Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation
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Incubation:

Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation
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Electron Microscopy:

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Protein Concentration:

Article Title: Cariogenicity induced by commercial carbonated beverages in an experimental biofilm-caries model
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    BioTek Instruments biofilms
    Biofilm formation. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose (BMGS), glucose (BMG) or sucrose (BMS). <t>Biofilms</t> were grown on polystyrene surface in 96 well plates and analyzed using a spectrophotometer. Results presented here represent mean absorbance at 575 nm (±standard deviation in error bars) from three independent experiments and ∗ and # P
    Biofilms, supplied by BioTek Instruments, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effects of chitosan-coated iron oxide nanoparticles on <t>biofilm</t> bacterial growth in the first 24 hours at the concentration of 500 µg/mL. Note: The results are expressed as mean ± SD (n = 3). Abbreviations: CFU, colony-forming unit; SD, standard deviation.
    Biofilm Biomass, supplied by BioTek Instruments, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biofilm formation. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose (BMGS), glucose (BMG) or sucrose (BMS). Biofilms were grown on polystyrene surface in 96 well plates and analyzed using a spectrophotometer. Results presented here represent mean absorbance at 575 nm (±standard deviation in error bars) from three independent experiments and ∗ and # P

    Journal: Frontiers in Microbiology

    Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

    doi: 10.3389/fmicb.2018.02130

    Figure Lengend Snippet: Biofilm formation. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose (BMGS), glucose (BMG) or sucrose (BMS). Biofilms were grown on polystyrene surface in 96 well plates and analyzed using a spectrophotometer. Results presented here represent mean absorbance at 575 nm (±standard deviation in error bars) from three independent experiments and ∗ and # P

    Article Snippet: After 24 h of incubation, the biofilms were stained in 0.1% crystal violet, bound dye was extracted using an acetone-ethanol mixture (1:4), and absorbance was measured using in Synergy II plate reader (BioTek) ( ).

    Techniques: Mutagenesis, Spectrophotometry, Standard Deviation

    Confocal microscopic analysis of biofilms. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose, glucose or sucrose. Biofilms were grown on HA disks vertically placed in 12 well plates for 24 h, and analyzed using a laser scanning confocal microscope. Panel shows representatives of the compressed confocal images at xy, yz, and xz axis of biofilms of UA159, TW416, and TW416C grown in BM plus glucose and sucrose.

    Journal: Frontiers in Microbiology

    Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

    doi: 10.3389/fmicb.2018.02130

    Figure Lengend Snippet: Confocal microscopic analysis of biofilms. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose, glucose or sucrose. Biofilms were grown on HA disks vertically placed in 12 well plates for 24 h, and analyzed using a laser scanning confocal microscope. Panel shows representatives of the compressed confocal images at xy, yz, and xz axis of biofilms of UA159, TW416, and TW416C grown in BM plus glucose and sucrose.

    Article Snippet: After 24 h of incubation, the biofilms were stained in 0.1% crystal violet, bound dye was extracted using an acetone-ethanol mixture (1:4), and absorbance was measured using in Synergy II plate reader (BioTek) ( ).

    Techniques: Mutagenesis, Microscopy

    SEM analysis of biofilms. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose, glucose or sucrose. Biofilms were grown on HA disks vertically placed in 12 well plates for 24 h, and analyzed using a scanning electron microscope (SEM). Panel shows images of UA159 and TW416 biofilms grown in BM plus glucose and sucrose, which were taken at magnification of 5,000 and 10,000 × g as indicated, with arrows indicating extracellular polymeric substances and asterisks indicating broken cells and cell debris, respectively.

    Journal: Frontiers in Microbiology

    Article Title: Deficiency of MecA in Streptococcus mutans Causes Major Defects in Cell Envelope Biogenesis, Cell Division, and Biofilm Formation

    doi: 10.3389/fmicb.2018.02130

    Figure Lengend Snippet: SEM analysis of biofilms. S. mutans wildtype (UA159), the mecA mutant (TW416) and its complement strain (TW416C) were grown in BM medium with glucose and sucrose, glucose or sucrose. Biofilms were grown on HA disks vertically placed in 12 well plates for 24 h, and analyzed using a scanning electron microscope (SEM). Panel shows images of UA159 and TW416 biofilms grown in BM plus glucose and sucrose, which were taken at magnification of 5,000 and 10,000 × g as indicated, with arrows indicating extracellular polymeric substances and asterisks indicating broken cells and cell debris, respectively.

    Article Snippet: After 24 h of incubation, the biofilms were stained in 0.1% crystal violet, bound dye was extracted using an acetone-ethanol mixture (1:4), and absorbance was measured using in Synergy II plate reader (BioTek) ( ).

    Techniques: Mutagenesis, Microscopy

    The activities of levofloxacin and Cath-A against the established biofilms of P. aeruginisa ( a ) and A . baumannii ( b ) at different concentrations (4–256 µg/mL). The antibiofilm effect of the peptide and levofloxacin was assessed by cv staining after 24 h of incubation. Biofilm formation was determined by measuring the absorbance at 595 nm using a microplate reader. Controls were untreated inoculated bacteria in Mueller Hinton (MH) broth. Data are shown as the mean ± SD of three independent tests. Significant different values were (# p

    Journal: Biomolecules

    Article Title: A Recombinant Snake Cathelicidin Derivative Peptide: Antibiofilm Properties and Expression in Escherichia coli

    doi: 10.3390/biom8040118

    Figure Lengend Snippet: The activities of levofloxacin and Cath-A against the established biofilms of P. aeruginisa ( a ) and A . baumannii ( b ) at different concentrations (4–256 µg/mL). The antibiofilm effect of the peptide and levofloxacin was assessed by cv staining after 24 h of incubation. Biofilm formation was determined by measuring the absorbance at 595 nm using a microplate reader. Controls were untreated inoculated bacteria in Mueller Hinton (MH) broth. Data are shown as the mean ± SD of three independent tests. Significant different values were (# p

    Article Snippet: The absorbance of the stained biofilm was measured at an optical density (OD) of 595 nm by a microplate reader (BioTek Instruments, PowerWave XS, Winooski, VT USA).

    Techniques: Staining, Incubation

    Images through field emission scanning electron microscopy (FE-SEM) (Magnification: 7000 X, Bar: 2 µm) of C. tropicalis biofilm; ( A ) control (1% DMSO) with densely clustered healthy cells; ( B ) amphotericin B (1 µg/mL); ( C ) citral (128 µg/mL); and ( D ) thymol (128 µg/mL), cells are more porous and flaccid with loosen cell surface. # Black arrows show pores in the cells.

    Journal: Journal of Fungi

    Article Title: In Vitro Anti-Biofilm Activities of Citral and Thymol Against Candida tropicalis

    doi: 10.3390/jof5010013

    Figure Lengend Snippet: Images through field emission scanning electron microscopy (FE-SEM) (Magnification: 7000 X, Bar: 2 µm) of C. tropicalis biofilm; ( A ) control (1% DMSO) with densely clustered healthy cells; ( B ) amphotericin B (1 µg/mL); ( C ) citral (128 µg/mL); and ( D ) thymol (128 µg/mL), cells are more porous and flaccid with loosen cell surface. # Black arrows show pores in the cells.

    Article Snippet: Briefly, the biofilm cells with/without treatment were added with 20 µM CM-H2DCFDA and incubated at 37 °C for 1 h. The fluorescence intensity was measured with excitation (485 nm) and emission (535 nm) using fluorescent microplate reader (BioTek, Synergy H1 Hybrid Multi-mode Microplate Reader, VT, USA).

    Techniques: Electron Microscopy

    ( A ) Confocal laser scanning microscopic images (CLSM) of C. tropicalis biofilm; dual stained: CON-A (Excitation: 488, Emission: 505), staining polysaccharide walls, green and FUN-1 (Excitation: 470, Emission: 590), staining metabolically active cells, red; thus dually staining yellowish-green colour of healthy biofilm; control (1% DMSO). Power field: 40×; Scale Bar: 50 µm; ( B ) COMSTAT analysis of C. tropicalis biofilm.

    Journal: Journal of Fungi

    Article Title: In Vitro Anti-Biofilm Activities of Citral and Thymol Against Candida tropicalis

    doi: 10.3390/jof5010013

    Figure Lengend Snippet: ( A ) Confocal laser scanning microscopic images (CLSM) of C. tropicalis biofilm; dual stained: CON-A (Excitation: 488, Emission: 505), staining polysaccharide walls, green and FUN-1 (Excitation: 470, Emission: 590), staining metabolically active cells, red; thus dually staining yellowish-green colour of healthy biofilm; control (1% DMSO). Power field: 40×; Scale Bar: 50 µm; ( B ) COMSTAT analysis of C. tropicalis biofilm.

    Article Snippet: Briefly, the biofilm cells with/without treatment were added with 20 µM CM-H2DCFDA and incubated at 37 °C for 1 h. The fluorescence intensity was measured with excitation (485 nm) and emission (535 nm) using fluorescent microplate reader (BioTek, Synergy H1 Hybrid Multi-mode Microplate Reader, VT, USA).

    Techniques: Confocal Laser Scanning Microscopy, Staining, Metabolic Labelling

    Graphical representation of the DCFDA fluorescence of C. tropicalis biofilm representing ROS production during treatment of amphotericin B, citral and thymol. Error bars denote standard deviation (SD).* p

    Journal: Journal of Fungi

    Article Title: In Vitro Anti-Biofilm Activities of Citral and Thymol Against Candida tropicalis

    doi: 10.3390/jof5010013

    Figure Lengend Snippet: Graphical representation of the DCFDA fluorescence of C. tropicalis biofilm representing ROS production during treatment of amphotericin B, citral and thymol. Error bars denote standard deviation (SD).* p

    Article Snippet: Briefly, the biofilm cells with/without treatment were added with 20 µM CM-H2DCFDA and incubated at 37 °C for 1 h. The fluorescence intensity was measured with excitation (485 nm) and emission (535 nm) using fluorescent microplate reader (BioTek, Synergy H1 Hybrid Multi-mode Microplate Reader, VT, USA).

    Techniques: Fluorescence, Standard Deviation

    Effects of chitosan-coated iron oxide nanoparticles on biofilm bacterial growth in the first 24 hours at the concentration of 500 µg/mL. Note: The results are expressed as mean ± SD (n = 3). Abbreviations: CFU, colony-forming unit; SD, standard deviation.

    Journal: International Journal of Nanomedicine

    Article Title: Reduced Staphylococcus aureus biofilm formation in the presence of chitosan-coated iron oxide nanoparticles

    doi: 10.2147/IJN.S41371

    Figure Lengend Snippet: Effects of chitosan-coated iron oxide nanoparticles on biofilm bacterial growth in the first 24 hours at the concentration of 500 µg/mL. Note: The results are expressed as mean ± SD (n = 3). Abbreviations: CFU, colony-forming unit; SD, standard deviation.

    Article Snippet: Biofilm biomass was quantified by the addition of 100 µL of 95% ethanol to each CV-stained microplate well, and the absorbance was determined with a plate reader (Bio-Tek, Winooski, VT, USA) at optical density (OD) 600.

    Techniques: Concentration Assay, Standard Deviation

    SEM images showing biofilms of the bacteria established on the surface of polystyren e plates. ( A ) In the absence of nanoparticles; ( B ) incubated with chitosan-coated iron oxide nanoparticles at 500 µg/mL; ( C ) incubated with chitosan-coated iron oxide nanoparticles at 4 mg/mL. Abbreviation: SEM, scanning electron microscopy.

    Journal: International Journal of Nanomedicine

    Article Title: Reduced Staphylococcus aureus biofilm formation in the presence of chitosan-coated iron oxide nanoparticles

    doi: 10.2147/IJN.S41371

    Figure Lengend Snippet: SEM images showing biofilms of the bacteria established on the surface of polystyren e plates. ( A ) In the absence of nanoparticles; ( B ) incubated with chitosan-coated iron oxide nanoparticles at 500 µg/mL; ( C ) incubated with chitosan-coated iron oxide nanoparticles at 4 mg/mL. Abbreviation: SEM, scanning electron microscopy.

    Article Snippet: Biofilm biomass was quantified by the addition of 100 µL of 95% ethanol to each CV-stained microplate well, and the absorbance was determined with a plate reader (Bio-Tek, Winooski, VT, USA) at optical density (OD) 600.

    Techniques: Incubation, Electron Microscopy

    Effects of different concentrations of nanoparticles on biofilm bacterial growth at after 24 hours of incubation. Notes: The results are expressed as mean ± SD (n = 3). * P

    Journal: International Journal of Nanomedicine

    Article Title: Reduced Staphylococcus aureus biofilm formation in the presence of chitosan-coated iron oxide nanoparticles

    doi: 10.2147/IJN.S41371

    Figure Lengend Snippet: Effects of different concentrations of nanoparticles on biofilm bacterial growth at after 24 hours of incubation. Notes: The results are expressed as mean ± SD (n = 3). * P

    Article Snippet: Biofilm biomass was quantified by the addition of 100 µL of 95% ethanol to each CV-stained microplate well, and the absorbance was determined with a plate reader (Bio-Tek, Winooski, VT, USA) at optical density (OD) 600.

    Techniques: Incubation

    Confocal images show the biofilm thickness after 48 hours of incubation. ( A ) In the absence of nanoparticles; ( B ) incubated with chitosan-coated iron oxide nanoparticles at a concentration of 500 µg/mL.

    Journal: International Journal of Nanomedicine

    Article Title: Reduced Staphylococcus aureus biofilm formation in the presence of chitosan-coated iron oxide nanoparticles

    doi: 10.2147/IJN.S41371

    Figure Lengend Snippet: Confocal images show the biofilm thickness after 48 hours of incubation. ( A ) In the absence of nanoparticles; ( B ) incubated with chitosan-coated iron oxide nanoparticles at a concentration of 500 µg/mL.

    Article Snippet: Biofilm biomass was quantified by the addition of 100 µL of 95% ethanol to each CV-stained microplate well, and the absorbance was determined with a plate reader (Bio-Tek, Winooski, VT, USA) at optical density (OD) 600.

    Techniques: Incubation, Concentration Assay