Fig. 4: Cocktails containing phages from three or more CG reliably eliminate multi-drug resistant clinical isolates of P. aeruginosa, including biofilm and polyclonal cultures in vivo.

a Growth curves for PA14 treated with phage(s) from DMS3vir, the same CG (DMS3vir + TIVP-H6), two different CGs (DMS3vir + OMKO1), or three different CGs (Luz24 + OMKO1 + PAML-31-1). Notably, the treatment with three phages from different CGs completely prevented growth (highlighted in yellow). All reported values are depicted from three independent experiments (n = 3) and associated error bars represent standard deviation from the mean. b Pooled Suppression Index data for PA14 treated with different phage cocktails; each data point represents as the average Suppression Index value of three independent experiments from a different combination of phages, with the number of phages and number of CGs as indicated. These cocktails are listed in Supplementary Table 2. The error bars represent standard deviation from the mean of pooled Suppression Index data. c Pooled Resistance Index for PA14 treated initially with different phage cocktails and subsequently re-challenged by each component phage that comprises the cocktails, all at an MOI of 100; each data point represents as the average Resistance Index value of three independent experiments. Detailed information on the phages used is in Supplementary Table 2. The error bars represent standard deviation from the mean of pooled Resistance Index data. d Representative three-dimensional renderings of PA14 biofilms after 72 hours in the absence or presence of phage therapy, as indicated. e Pooled data for PA14 cells in biofilms treated with different phage cocktails; each data point represents as the average cell number of three independent experiments. The error bars represent standard deviation from the mean of pooled biofilm data. f Suppression Index for 16 clinical isolates of P. aeruginosa by individual phages Luz24, OMKO1, and PAML-31-1, each at an MOI of 100, or a cocktail of these three phages (KIM-C1) with a combined MOI of 100. g, Suppression Index for these clinical isolates of P. aeruginosa by cocktail KIM-C1 at combined MOIs of 1, 100, or 500. Data in f and g are shown as an average of triplicate independent results. h Suppression Index for monoclonal and polyclonal P. aeruginosa cultures treated individually with KIM-C1 cocktail, its three constituent phages, or KIM-M1 mocktail (a mixture of 3 phages with same CG: DMS3vir, KOR_P1, and KOR_P2) grown in planktonic form or as biofilms. Data are shown as an average of triplicate independent results. The phage dose used here is a combined MOI of 100. i A schematic for wound mice model. This schematic model was created in BioRender. Kim, K. (2023) BioRender.com/s43l916. j Representative wound images of each condition (control, cocktail, and mocktail) over the course of experiments. k CFU of the polyclonal cultures in wounds under the conditions in j. The red line indicates the average value of each condition from at least 15 independent experiments. The black dotted line with “LOD” denotes the limit of detection, which is 100 CFU/g (log(100) = 2). The data under LOD indicates LOD/2 (50 CFU/g or log(50) = 1.7). Statistical significance in b, c, e, and k is derived from One-way analysis of variance (ANOVA) and significance levels include *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Source data are provided as a Source Data file.