Drug-drug interaction using checkerboard assay. Drug interaction was evaluated in MIC₅₀, one-half the MIC₅₀, one-quarter the MIC₅₀, one half of one quarter the MIC₅₀ of CLA (horizontal) in combination with MIC₅₀, one-half the MIC, one-quarter the MIC₅₀, one half of one quarter the MIC₅₀ of OMD (A), AMK (C), and RFB (E). Isobolograms of the resazurin checkerboard synergy testing method showing synergy of CLA with OMD (B). The additive effect observed when CLA interact with AMK (D) and RFB (F). The white line indicates MIC₅₀ value of each compound.

Estimation of bactericidal effect by CFU counts. Mab was grown in the presence of different concentrations of CLA alone or in combination with decreasing concentrations of OMD (A) and AMK (B). Following 7 days of culture, Mab were plated to 7H10 agar plate to determine live bacteria. The DMSO treated bacteria were also plated on day 0 and on day 7. One-way ANOVA with Tukey?s multiple comparison test was used to compare the means across multiple groups (**p < 0.01; ***p < 0.001).

ZF in vivo efficacy of CLA-OMD. The drug concentrations that show 1 log₁₀ CFU reduction were determined using different concentrations of CLA, BDQ, AMK, OMD, RFB, and CFX in Mab infected ZF model (A). To determine in vivo efficacy, survival curve was plotted from MabR-mWasabi infected ZF for 13 days (n = 20, representative of three independent experiments) (B). Each different combination treatment was carried out. CLA (3.1 ?M) was combined with BDQ (3.1 ?M), AMK (12.5 ?M), OMD (6.3 ?M), RFB (6.3 ?M), and CFX (6.3 ?M) respectively. Survival curves were constructed using the log-rank (Mantel-Cox) test (**p < 0.01; ***p < 0.001). Inf UNT: Infected but not treated control. Therapeutic outcome using drug combinations was validated by traditional agar plate quantification method (C). Data was expressed as the mean log₁₀ CFU per embryo (n = 10 of each condition) from three independent experiments. Drug combination effect was also observed using fluorescence under microscope. Each drug combinations were treated to the ZF infected with MabR-mWasabi and reduction of mWasabi signal in ZF was monitored under the fluorescent microscope (D).