Fig. 1

Effect of zebrafish biparental exposure to difenoconazole on the development of F1 generation embryos/larvae. (A) Number of spontaneous movement in 20 s of embryos at 24 hpf; (B) heartbeat of 48 hpf embryos within 20 s; (C) malformation rate of 48 hpf embryos; (D) hatching rate of F1 generation at 72 hpf and 96 hpf; (E) mortality of F1 generation at 24 hpf and 96 hpf; (F) body length of F1 generation at 96 hpf. (*p < 0.05; **p < 0.01).

Fig. 2

Transcriptomic alterations in F1 generation larvae. (A) Volcano plot of DEGs in F1 generation embryos of zebrafish parents exposed to 0.1 ?g/L difenoconazole; (B) volcano plot of DEGs in F1 generation embryos of zebrafish parents exposed to 1.0 ?g/L difenoconazole. (C) Compared with control, KEGG enrichment analysis of DEGs in the 0.1 ?g/L difenoconazole group. (D) Compared with control, KEGG enrichment analysis of DEGs in the 1.0 ?g/L difenoconazole group.

Fig. 3

Activities of enzymes related to apoptosis, autophagy, and energy metabolism in F1 generation larvae after biparental exposure to difenoconazole. (A) Caspase3 activity, (B) Caspase9 activity, (C) CATD activity, (D) PK activity, and (E) ACTA activity. (*p < 0.05; **p < 0.01).

Fig. 4

Correlation analysis between F1 generation developmental indices and enzyme activities related to apoptosis, autophagy, and energy metabolism in F1 generation larvae. Correlation is significant at the 0.05 level (*p < 0.05; **p < 0.01).

Fig. 5

Effect of F0 generation female or male fish exposure to difenoconazole on the development of the hybrid F1 generation embryos/larvae. (A) Number of spontaneous movement in 20 s of embryos at 24 hpf; (B) heartbeat of 48 hpf embryos within 20 s; (C) malformation rate of 48 hpf embryos; (D) hatching rate of F1 generation at 72 hpf; (E) body length of F1 generation at 120 hpf; (F) mortality of F1 generation at 24 hpf and 120 hpf. Female represents F1 generation embryos/larvae produced by the pairing of difenoconazole-exposed F0 generation females and nondifenoconazole-exposed males; male represents the F1 generation embryos/larvae produced by the pairing of difenoconazole-exposed F0 males and nondifenoconazole-exposed females. (*p < 0.05; **p < 0.01).

Fig. 6

Activities of enzymes related to apoptosis, autophagy, and energy metabolism in the hybrid F1 generation larvae after F0 generation female or male fish exposure to difenoconazole. (A) Caspase3 activity, (B) Caspase9 activity, (C) CATD activity, (D) PK activity, and (E) ACTA activity. Female represents the FF1; Male represents the MF1. (*p < 0.05; **p < 0.01).

Fig. Abs

Acknowledgments
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