Fig. 6: Antibiofilm and antimicrobial activity of the phage cocktail. | Nature Communications

Fig. 6: Antibiofilm and antimicrobial activity of the phage cocktail.

From: Targeting Pseudomonas aeruginosa biofilm with an evolutionary trained bacteriophage cocktail exploiting phage resistance trade-offs

Fig. 6: Antibiofilm and antimicrobial activity of the phage cocktail.

a Efficiency of plating (EOP) of five phages (FJK, FJK.R9-15, FJK.R9-30, MK, and MK.R3-15) on 18 Paer09 strains treated with the phage cocktail (FJK.R9-30 + MK.R3-15). EOP is defined within the methods. Mutation-associated protein of each strain with amino acid change in parentheses (1c.203_204dupTC p.R69fs and 2c.205delA p.R69fs). Lack of labelling refers to isolates with no apparent mutation. b Illustration of the number of isolates by mutant protein and treatment group (FJK; FJK.R9-15; FJK.R9-30; Cocktail). c qPCR-determined bacterial load (CFU/bead) of 24 h pre-established Paer09 biofilm untreated (GC) and after co-incubation (3, 6 and 24 h) with FJK.R9-30 or the cocktail. The p-value is 0.0077 (6 h, Cocktail vs. FJK.R9-30). d, e Heat flow (µW) and heat (J) curves measured for 48, 72 and 96 h of Paer09 biofilm co-incubated with MK.R3-15, FJK.R9-30, cocktail (individual replicates) or untreated (GC, dashed black line). Minimum heat flow values in the right upper corner (d). The p-values are 0.0404 (FJK.R9-30 vs. Cocktail) and 0.0024 (MK.R3-15 vs. Cocktail). f Calculated lag time (h) from the heat curves for each tested condition. The p-values are 0.0051 (FJK.R9-30 vs. Cocktail) and 0.0002 (MK.R3-15 vs. Cocktail). OD600 measurement of a seven-day (g) and three-day (h) co-incubation of planktonic naive Paer09 strain with the cocktail (MOI: 0.001, g; MOI: 0.0001, h). Thin lines represent the individual replicates (n = 8), and the thick coloured line the average, excluding completely suppressed replicates (n = 4, g; n = 2, h). i Illustration of averaged co-incubation curves (including grey, violet, and yellow curves from Fig. 5d) over three days compared to a growth control (dotted black line) and negative control (black line). The cocktail (MOI: 0.0001) is shown as a dotted green curve. All experiments were performed in fourfold and unless specified otherwise, the calorimetric curves (d, e) display the mean. The error bars represent the standard error of the mean. Statistical analysis was conducted by unpaired two-tailed Student’s t test and p-values indicated with asterisks (*p = <0.05; **p = <0.01; ***p = <0.001). Source data are provided as a source data file.

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