毒死蜱和百菌清对斑马鱼早期发育阶段的联合毒性效应

Combined effects of chlorpyrifos and chlorothalonil on early development stage of zebrafish

为评价毒死蜱和百菌清的单剂毒性和联合毒性,以斑马鱼为受试生物,采用静态法和实时荧光定量PCR方法,研究毒死蜱和百菌清单剂,以及二元组合时对斑马鱼胚胎的急性毒性和对斑马鱼幼鱼性腺轴相关基因的影响。结果显示,毒死蜱和百菌清对斑马鱼胚胎分别表现为中等和高等毒性,96 h-LC_(50)(半致死浓度)分别为2.139 mg·L^(-1)和0.353 mg·L^(-1)。毒死蜱和百菌清联合作用类型为协同作用,联合暴露时,对斑马鱼胚胎的96 h-LC_(50)分别为0.38 mg·L^(-1)和0.063 mg·L^(-1)。中浓度(26.7μg·L^(-1))和高浓度(106.7μg·L^(-1))的毒死蜱可以诱导斑马鱼幼鱼体内芳香化酶基因(CYP19α)、卵黄蛋白原基因(VTG1、VTG2)和雌激素受体基因(ERα、ERβ1、ERβ2)不同程度的上调表达;低浓度(1.1μg·L^(-1))百菌清诱导斑马鱼体内VTG1、VTG2、ERβ1和ERβ2基因的上调表达;低浓度(6.7μg·L^(-1)毒死蜱+1.1μg·L^(-1)百菌清)二元组合会引起斑马鱼体内VTG1、ERβ1、ERβ2和ERα基因下调。说明毒死蜱和百菌清对斑马鱼幼鱼具有雌激素内分泌干扰效应,但毒死蜱和百菌清二元组合对斑马鱼幼鱼的雌激素内分泌干扰效应降低。在评估农药对水生生物的危害时,应充分考虑复合污染时的联合效应。

To evaluate the toxicity of chlorpyrifos and chlorothalonil in single dose and combinations,zebrafish was taken as biological subjects.By using the static method and real-time fluorescent quantitative PCR,the acute toxicity of chlorpyrifos or/and chlorothalonil on zebrafish embryos and their influence on the related genes of zebrafish larvae gonadal axis were studied.It was shown that chlorpyrifos and chlorothalonil were moderately and highly toxic to zebrafish embryos,respectively,as the 96 h-LC_（50）（lethal concentration 50）was 2.139 mg·L-1and 0.353 mg·L-1,respectively.The toxicivy of chlorpyrifos and chlorothalonil were synergistic to zebrafish embryos.In the joint exposure of chlorpyrifos and chlorothalonil,the 96 h-LC_（50）was 0.38 mg·L-1and 0.063 mg·L-1,respectively.Moderate concentration（26.7μg·L-1）and high concentration（106.7μg·L-1）of chlorpyrifos led to upregulation of the gene expression of the aromatase（CYP19α）,vitellogenin（VTG1,VTG2）,and estrogen receptor（ERα,ERβ1 and ERβ2）.Low concentration of 1.1μg·L-1chlorothalonil led to the upregulation of gene expression of VTG1,VTG2,ERβ1 and ERβ2.Combination of 6.7μg·L-1chlorpyrifos＋1.1μg·L-1chlorothalonil led to the downregulation of gene expression of VTG1,ERβ1,ERβ2 and ERαin zebrafish compared with the control.The results indicated that chlorpyrifos and chlorothalonil had an estrogenic effect on zebrafish,while the combination reduced the effect of estrogenic interference.Therefore,the effects of combined pollution should be fully considered for harm assessment of pesticides to aquatic organisms.