Trifloxystrobin (TRI) is a methacrylate fungicide, and fluopyram (FLU) is a new pyridylethylbenzamide fungicide and nematicide. Both are often detected in water bodies and may be highly toxic to many aquatic organisms. Unfortunately, the aquatic biological risks of single FLU or a mixture of trifloxystrobin and fluopyram have not been reported. In this study, zebrafish was selected as the test organism to investigate the combined toxicity of trifloxystrobin and fluopyram to zebrafish. After zebrafish embryos exposed to three pesticide solutions, Alcian-blue staining, Alizarin-red staining and quantitative PCR (qPCR) were performed. The results indicated that 96h-LC₅₀ of TRI was 0.159 mg·L⁻¹ to zebrafish embryo, which was highly toxic. The 96h-LC₅₀ of FLU to zebrafish embryos was 4.375 mg·L⁻¹, being moderately toxic. The joint toxicity to zebrafish embryos(FLU at 96h-LC₅₀ and TRI at 96h-LC₅₀ in a 1:1 weight ratio to form a series of concentration treatment groups) was antagonistic. Both trifloxystrobin and fluopyram also inhibited the skeletal development of zebrafish and showed to be antagonistic. The results of qPCR indicated upregulations of different genes upon three different treatments. TRI mainly induced Smads up-expression, which may affect the BMP-smads pathway. FLU mainly induced an up-expression of extracellular BMP ligands and type I receptor (Bmpr-1a), which may affect the BMP ligand receptor pathway. The 1:1 mixture (weight ratio) of trifloxystrobin and fluopyram induced a reduction of the genes of extracellular BMP ligand (Smads) and type I receptor (Bmpr1ba), which may down-regulate BMP signaling and thus attenuating cartilage hyperproliferation, hypertrophy and mineralization. The results warren an interest in further studying the effect of the two fungicides in a mixture on zebrafish.

肟菌酯 (TRI) 是一种甲基丙烯酸酯类杀菌剂，氟吡菌酰胺 (FLU) 是一种新型吡啶乙基苯甲酰胺杀菌剂和杀线虫剂。两者经常在水体中被发现，并且可能对许多水生生物具有剧毒。不幸的是，单一FLU或肟菌酯和氟吡菌酰胺混合物的水生生物风险尚未有报道。本研究以斑马鱼为试验生物，考察肟菌酯和氟吡菌酰胺对斑马鱼的联合毒性。将斑马鱼胚胎暴露于三种农药溶液后，进行阿新蓝染色、茜素红染色和定量 PCR (qPCR)。结果表明，TRI对斑马鱼胚胎的96h-LC ₅₀为0.159 mg·L ^-1，具有剧毒。FLU对斑马鱼胚胎的96h-LC ₅₀为4.375 mg·L ^-1，具有中等毒性。对斑马鱼胚胎的联合毒性（FLU 96h-LC ₅₀和TRI 96h-LC ₅₀按1:1重量比组成系列浓度处理组）呈拮抗作用。肟菌酯和氟吡菌酰胺均对斑马鱼骨骼发育有抑制作用，并表现出拮抗作用。qPCR 结果表明三种不同处理后不同基因的上调。TRI主要诱导Smads上表达，这可能影响BMP-smads通路。FLU主要诱导细胞外BMP配体和I型受体( Bmpr-1a )的上调表达，这可能影响BMP配体受体通路。肟菌酯和氟吡菌酰胺的 1:1 混合物（重量比）可诱导细胞外BMP配体 (Smads) 和 I 型受体 ( Bmpr1ba )基因的减少，从而下调BMP信号传导，从而减弱软骨过度增殖、肥大和矿化。这一结果引起了人们对进一步研究两种杀菌剂混合物对斑马鱼的影响的兴趣。