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Fig. 1 Experimental design for the study. (A) shows the design to test whether there was an effect of BaP and the gut microbiome on behavior in zebrafish. Embryos were exposed to three different concentrations (noted by the colors) in 96-well plates as shown, then measured via EPR, allowed to grow to the larval stage, and LPR behavior assays. Larvae were subsequently dissected for gut microbiome data. (B) shows the study design for the effect of gut microbiome presence on the relationship between BaP and behavior. Similarly, colors represent a gradient of BaP exposures that were applied to gnotobiotic, CV, and CVZ zebrafish embryos in 96-well plates and EPR, LPR, and microbiome data collected. This study design allows us to determine not only the associations between BaP, gut microbiome, and behavior, but also the combined effect of BaP and gut microbiome state on behavior.