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采用改进的ELLMAN法测定凡纳滨对虾(Litopenaeusvannamei)脑神经节乙酰胆碱酯酶(acetylcholinesterase,AChE)活性,确定其适宜测定条件,并在此基础上测定对虾不同组织AChE活性,比较毒死蜱、敌敌畏、辛硫磷和乙酰甲胺磷4种有机磷农药对凡纳滨对虾脑神经节AChE活性的影响。结果表明,温度35℃,磷酸缓冲液pH 7.5时,凡纳滨对虾的AChE活性最高,保温时间对酶活性影响很小;对虾AChE活性存在着明显的组织分布差异性,脑神经节AChE活性最高,为(49.73±8.42)nmol/(min.mg),分别是鳃、肌肉和肝胰腺的3倍、15倍和19倍;对虾AChE对敌敌畏最为敏感,IC50为0.19μg/mL,对毒死蜱和辛硫磷敏感性较强,IC50分别为7.20μg/mL和9.39μg/mL,AChE对乙酰甲胺磷敏感性最弱,IC50为136.77μg/mL。由此可见,对虾养殖中应注意防范敌敌畏、毒死蜱和辛硫磷等有机磷农药的毒性危害。研究亮点:有机磷农药是常用杀虫剂,其残留危害着水产养殖,以前对水产动物毒性研究主要集中在鱼类,很少关注对有机磷农药尤为敏感的对虾。本文优化建立了对虾AChE活性检测方法,评价分析了4种有机磷农药对对虾AChE活性的抑制作用,为今后开展对虾有机磷毒理学研究奠定了基础,也为健康养殖提供了技术支持。
The activity of acetylcholinesterase (AChE) in cranial ganglion of Litopenaeus vannamei was determined by a modified ELLMAN method. The suitable assay conditions were determined. AChE activity in different tissues of shrimp was determined. Chlorpyrifos, dichlorvos, Effects of 4 kinds of organophosphorus pesticides including parathion, acetamidine and methyl thioacetate on AChE activity in cranial ganglion of. The results showed that the AChE activity of Litopenaeus vannamei was the highest at 35 ℃ and pH 7.5, and the incubation time had little effect on the enzyme activity. AChE activity of shrimp showed obvious tissue distribution, and AChE activity of cranial ganglia was the highest (49.73 ± 8.42) nmol / (min.mg), respectively, which were 3 times, 15 times and 19 times higher than that of gills, muscle and hepatopancreas. The AChE of shrimp was the most sensitive to dichlorvos with IC50 of 0.19μg / mL, Phoxim was more sensitive with IC50 of 7.20μg / mL and 9.39μg / mL, respectively, and AChE was the weakest for acephate with IC50 of 136.77μg / mL. Thus, shrimp farming should pay attention to prevent dichlorvos, chlorpyrifos and phoxim and other organophosphate pesticides toxicity hazards. Research highlights: Organophosphorus pesticides are commonly used pesticides, the residues endanger aquaculture, aquatic animal toxicity previously focused on fish, with little concern for organophosphate pesticides are particularly sensitive to shrimp. This paper optimized the detection of AChE activity in shrimp, evaluated and evaluated the inhibition of AChE activity of four organophosphorus pesticides on shrimp, laid the foundation for the future research on organophosphate toxicology of shrimp and provided technical support for healthy breeding.