Developmental toxicity and inflammatory response induced by Botrytis cinerea in zebrafish (Danio rerio) larvae

Botrytis cinerea can reduce the yield of fruits and vegetables by infecting plants. The conidia produced by Botrytis cinerea can be transmitted to the aquatic environment via air and water, but the effects of Botrytis cinerea on aquatic animals is unknown. In this research, the influence of Botrytis cinerea on the development, inflammation, and apoptosis of zebrafish larvae and the underlying mechanism was evaluated. Results indicated that, compared with the control group, the larvae exposed to 10¹-10³ CFU/mL of Botrytis cinerea spore suspension had a delayed hatching rate, lower head and eye area, shorter body length, and larger yolk sac at 72 h post-fertilization. In addition, the quantitative fluorescence intensity of treated larvae displayed a dose-dependent increase in apoptosis sign, revealing that Botrytis cinerea could generate apoptosis. Subsequently, zebrafish larvae were inflamed after exposure to Botrytis cinerea spore suspension, which was characterized as inflammatory infiltration and macrophage aggregation in the intestine. The enrichment of the pro-inflammatory factor TNF-α activated the NF-κB signaling pathway, generating the increase of the transcription level of target genes (jak3, pi3k, pdk1, akt, and ikk2) and the high expression of major proteins NF-κB (P65) in this pathway. Likewise, elevated content of TNF-α could activate JNK, which turned on the P53 apoptotic pathway, leading to a significant increase in the bax, caspase3, and caspase9 transcript abundances. This study demonstrated that Botrytis cinerea could cause developmental toxicity, morphological malformation, inflammation, and cell apoptosis in zebrafish larvae, which provided data support and a theoretical basis for ecological health risk assessment and filled the gap in biological research of Botrytis cinerea.