In the present study OMICs analysis was employed to investigate the early molecular responses of zebrafish embryos to exposure to the fungicide difenoconazole. Difenoconazole, a sterol biosynthesis inhibitor according to Fungicide Resistance Action Committee (FRAC) classification, may also induce adverse effects on non-target organisms inhabiting the environment. Early molecular responses in terms of transcriptome and proteome analysis were investigated and refined to select potentially substance specific biomarker candidates for early prediction of difenoconazole toxicity in zebrafish embryos.