The toxicity of β-diketone antibiotics(DKAs)to larval and adult zebrafish(Danio rerio)was investigated by miRNA sequencing and bioinformatics analyses.In control and DKA-exposed groups,215 differentially expressed miRNAs were screened,and 4076 differential target genes were predicted.Among 51 co-differentially expressed genes,45 were annotated in KOG functional classification,and 34 in KEGG pathway analysis.The homology analysis of 20 miRNAs with human hsa-miRNAs demonstrated 17 high homologous sequences.The expression levels of 12 miRNAs by qRT-PCR were consistent with those by sRNA-seq.A regulatory network for 4 positive miRNA genes(dre-miR-10,-96,-92 and-184)was plotted,and the high-degree of connectivity between miRNA-gene pairs suggests that these miRNAs play critical roles during zebrafish development.The consistent expression of dre-miR-184 and dre-miR-96 was proved in 120-hpf zebrafish brain,gill,otoliths and lateral line neuromast by qRT-PCR,miRNA-seq,W-ISH and ISH.DKA-exposure led to vacuolation of interstitial cells,reduced number of neurons,glial cell proliferation and formation of glial scar,and the obvious abnormality of cell structure might result from abnormal expression of differentially expressed miRNAs.In general,chronic DKA-exposure resulted in comprehensively toxic effects on larval and adult zebrafish tissues,especially for nervous system.