新烟碱类杀虫剂对非靶标生物的潜在毒性影响已经引起了国内外的普遍关注，但是有关该类杀虫剂在低剂量长周期暴露下对土壤生物的毒性效应及其机理的研究十分薄弱。本项目拟选择六种新烟碱类杀虫剂（吡虫啉、啶虫脒、噻虫胺、烯啶虫胺、噻虫啉和呋虫胺等）为对象，以土壤无脊椎动物蚯蚓为受试生物。根据其对蚯蚓急性和亚急性毒性指标（体重、繁殖及回避行为等）影响的毒性信息，采用单细胞凝胶电泳、液相色谱、蛋白质印迹分析和逆转录PCR等技术，探究对子代蚯蚓形态、体重及生化酶活性、DNA损伤、基因差异表达等的影响，明确其变化规律，并与亲代蚯蚓进行比较，绘制出变化的动态曲线，以期阐明新烟碱类杀虫剂对蚯蚓的潜在毒性风险及其致毒机理，并筛选出用于早期诊断土壤低剂量农药污染的敏感生物标志物。本项目研究结果可为低剂量农药长期暴露污染土壤生态毒理诊断提供方法论，同时亦为农药合理使用和生态环境保护提供基础科学依据。

The potential toxicity of neonicotinoid insecticides to non-target organisms has attracted widespread attention, but little has been done regarding the toxicity and its mechanism of the insecticides to soil organisms following long-term and low-dose exposure. In this study, six neonicotinoid insecticides (imidacloprid, acetamiprid, clothianidin, nitenpyram, thiacloprid and dinotefuran) were investigated for their acute and subacute toxicity to the soil invertebrate earthworm based on body weight, reproduction and avoidance behavior. Using single cell gel electrophoresis, liquid chromatography, Western blot analysis and reverse transcription PCR technology, we studied and analyzed the impact of the insecticides on the morphology, body weight, enzyme activity, DNA damage and gene expression in earthworm offsprings. The changes between the parental and offspring earthworms were compared to draw the dynamic curve of change in order to elucidate the potential toxicity risks of neonicotinoid insecticides on earthworm and its toxicity mechanism. These data were used to screen sensitive biomarkers for early diagnosis of low-dose pesticide pollution of soil. The results of this work can provide diagnostic methods for pesticide soil pollution following long-term and low-dose exposure, and scientific basis of rational pesticide use and protection of ecological environment.