Journal: NPJ Biofilms and Microbiomes

Article Title: Biofilm dynamics: linking in situ biofilm biomass and metabolic activity measurements in real-time under continuous flow conditions

doi: 10.1038/s41522-020-00153-9

Figure Lengend Snippet: a PA01 biofilms were cultivated in the combined CEMS-BioSpec system with 10 g L −1 TSB supplied at a flow rate of 12.5 mL h −1 . Changes in biofilm metabolic activity (black line) and biofilm biomass (magenta line) were monitored in real-time over the course of 125 h. Total protein from biofilm biomass (red dashed line) was sampled eight times during the first 75 h, each point is the mean of biological duplicates, technical triplicates with the standard deviation indicated by the error bar ( n = 6). b Direct correlation of total protein isolated from the sessile biomass with the corresponding BioSpec absorbance measurements of attached biomass ( r = 0.96, r 2 = 0.92, p < 0.05). Simple linear correlation was performed with Pearson correlation coefficient ( r ) and coefficient of determination ( R 2 ) calculated for each correlation.

Article Snippet: BioSpec was combined with the previously developed carbon dioxide evolution measurement system (CEMS) to allow simultaneous measurement of biofilm biomass and metabolic activity and revealed a differential response of these interrelated parameters to changing environmental conditions.

Techniques: Activity Assay, Standard Deviation, Isolation

Journal: NPJ Biofilms and Microbiomes

Article Title: Biofilm dynamics: linking in situ biofilm biomass and metabolic activity measurements in real-time under continuous flow conditions

doi: 10.1038/s41522-020-00153-9

Figure Lengend Snippet: P. aeruginosa PAO1 biofilms were cultivated under flow with a nutrient concentration of 3 g L −1 TSB until whole-biofilm metabolic activity (black line) and approximate biomass (magenta line) steady-state was reached (0 ± 70 h) ( a ). The inflow of nutrient solution was replaced with a solution of diluted sodium hypochlorite diluted with dH 2 O (1:100, 570 ppm free chlorine). The sodium hypochlorite solution was aseptically introduced into the system using the same flow rate for a treatment period of 1 h (green shaded region) followed by reintroduction of nutrient medium (recovery phase, post 71 h). b Zoomed-in view showing a 15 h period spanning pre-perturbation and recovery phase. The free chlorine concentrations in the effluent from the CEMS-BioSpec system (red line) was determine prior to the start of the treatment period, during and for 1 h after the treatment period (±70–71 h). Error bars of all free chlorine data points represent the standard error of triplicate readings at each time point ( n = 3).

Article Snippet: BioSpec was combined with the previously developed carbon dioxide evolution measurement system (CEMS) to allow simultaneous measurement of biofilm biomass and metabolic activity and revealed a differential response of these interrelated parameters to changing environmental conditions.

Techniques: Concentration Assay, Activity Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: Biofilm dynamics: linking in situ biofilm biomass and metabolic activity measurements in real-time under continuous flow conditions

doi: 10.1038/s41522-020-00153-9

Figure Lengend Snippet: Culture media and test fluids were introduced via a manifold ( a ) from the respective reservoirs with a peristaltic ( b ) to the respective systems contained in a heating/cooling water bath (26 °C). The CO 2 -Free sweeper gas was introduced into the annular space of CEMS (Expanded B, red shaded region) via a gas mass flow controller, allowing for the collection of biofilm-evolved CO 2 and analyzed by an infrared CO 2 analyzer. Biofilm biomass was measured between the two CEMS units (expanded B) via the internal silicon tube (containing biofilm biomass) being passed through a cavity with a LED illuminating the tube from one side and a digital light sensor on the transverse side measuring the amount of illumination absorbed by biomass in the tube. All dimensions are to the nearest mm.

Article Snippet: BioSpec was combined with the previously developed carbon dioxide evolution measurement system (CEMS) to allow simultaneous measurement of biofilm biomass and metabolic activity and revealed a differential response of these interrelated parameters to changing environmental conditions.

Techniques:

Journal: NPJ Biofilms and Microbiomes

Article Title: Biofilm dynamics: linking in situ biofilm biomass and metabolic activity measurements in real-time under continuous flow conditions

doi: 10.1038/s41522-020-00153-9

Figure Lengend Snippet: a PA01 biofilms were cultivated in the combined CEMS-BioSpec system with 10 g L −1 TSB supplied at a flow rate of 12.5 mL h −1 . Changes in biofilm metabolic activity (black line) and biofilm biomass (magenta line) were monitored in real-time over the course of 125 h. Total protein from biofilm biomass (red dashed line) was sampled eight times during the first 75 h, each point is the mean of biological duplicates, technical triplicates with the standard deviation indicated by the error bar ( n = 6). b Direct correlation of total protein isolated from the sessile biomass with the corresponding BioSpec absorbance measurements of attached biomass ( r = 0.96, r 2 = 0.92, p < 0.05). Simple linear correlation was performed with Pearson correlation coefficient ( r ) and coefficient of determination ( R 2 ) calculated for each correlation.

Article Snippet: Sessile growth dynamics were studied under continuous flow conditions within the CEMS linked to the BioSpec, allowing for the simultaneous real time, nondestructive monitoring of biofilm metabolism, and biomass respectively (Fig. ).

Techniques: Activity Assay, Standard Deviation, Isolation

Journal: NPJ Biofilms and Microbiomes

Article Title: Biofilm dynamics: linking in situ biofilm biomass and metabolic activity measurements in real-time under continuous flow conditions

doi: 10.1038/s41522-020-00153-9

Figure Lengend Snippet: P. aeruginosa PAO1 biofilms were cultivated under flow with a nutrient concentration of 3 g L −1 TSB until whole-biofilm metabolic activity (black line) and approximate biomass (magenta line) steady-state was reached (0 ± 70 h) ( a ). The inflow of nutrient solution was replaced with a solution of diluted sodium hypochlorite diluted with dH 2 O (1:100, 570 ppm free chlorine). The sodium hypochlorite solution was aseptically introduced into the system using the same flow rate for a treatment period of 1 h (green shaded region) followed by reintroduction of nutrient medium (recovery phase, post 71 h). b Zoomed-in view showing a 15 h period spanning pre-perturbation and recovery phase. The free chlorine concentrations in the effluent from the CEMS-BioSpec system (red line) was determine prior to the start of the treatment period, during and for 1 h after the treatment period (±70–71 h). Error bars of all free chlorine data points represent the standard error of triplicate readings at each time point ( n = 3).

Article Snippet: Sessile growth dynamics were studied under continuous flow conditions within the CEMS linked to the BioSpec, allowing for the simultaneous real time, nondestructive monitoring of biofilm metabolism, and biomass respectively (Fig. ).

Techniques: Concentration Assay, Activity Assay