Fig. 1

Schematic representation of the experimental procedure: exposure period in 2 generation, endpoints analyzed in each sampling point. Abbreviation: EX represents that F1 generation was exposed to CBZ in 3 treatment groups and a control group as their parental exposure procedure; NE represents that F1 generation was placed in CBZ-free water for recovery.

Fig. 2

A, Cumulative spawning in F0 zebrafish exposed to various concentrations of CBZ. The data represents the mean cumulative number of eggs per female collected from different exposed concentrations. B, Relative abundance of the top 10 phyla in F0. C, Relative abundance of the top 10 species in F0. Histological changes in adult zebrafish (F0) after 21 days exposure to CBZ: (D) Control; (E) 1 μg/l; (F) 10 μg/l; (G) 100 μg/l. The black arrow is hepatocyte and hepatocyte cytoplasmic loosening or vacuolization. The scale bar is 50 μm. H, Contents of IL-1β, IL-8, TNF-α, and LYS in F0 generation (*—p < .05; **—p < .01; ***—p < .001; ****—p < .0001).

Fig. 3

Effects and proposed toxicity pathway on the innate immune and adaptive immune system in F1 generation at 5 and 21 dpf. It showed the effects in different biological levels in adult zebrafish (F0) exposed to 3 CBZ treatments and the transcription level of genes in F1 which exposed or nonexposed to CBZ (A), and the effects in F1 exposed to CBZ added LPS or not added LPS (B). The color key (red, green, gray) indicates significant upregulation, downregulation, and no significant difference compared with the control group

Fig. 4

Cytokines (IL-1β, IL-8, TNF-α) and LYS content of F1 (from parental generation exposed to 3 treatments and the control group) continued to be exposed to CBZ (1, 10, and 100 μg/l) or nonexposed to CBZ at 5 dpf.

Fig. 5

Respiratory burst activity of PMA-stimulated F1 zebrafish continuously exposed or nonexposed to CBZ (group I)

Fig. 6

Movement of F1 zebrafish following parental exposure to the control group and treatments (1, 10, and 100 μg CBZ/L) at 5 dpf. A, Movement of F1 larvae exposed to CBZ (group I). B, Movement of F1 larva exposed to CBZ added LPS (group III)

Fig. 7

Proposed toxicity pathway for a mechanistic understanding of carbamazepine-mediated immunological effects across 2 generations in zebrafish

Acknowledgments
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