鸭胚性腺组织微囊藻毒素检测方法建立要点分析.doc

鸭胚性腺组织微囊藻毒素检测方法建立 摘要：微囊藻毒素（Microcystins）是一类具有生物活性的七肽类毒素，由淡水中的蓝藻细菌生产。由于现代工业产业的快速发展，使得水体污染日益严重，富营养化现象加剧，不断繁殖的蓝藻细菌所产生的大量的微囊藻毒素已经严重危害到生活在其周围环境中的动物以及人类的健康。本实验通过对注射了适当剂量纯微囊藻毒素的鸭胚性腺进行酶联免疫吸附试验，检测不同孵化天数下，性腺中富集的微囊藻毒素的量。结果表明，纯种的微囊藻毒素会对鸭胚胎的生长发育造成一定影响，并在性腺组织中富集，且大于0.1μg/egg的剂量将会导致鸭胚大量死亡。 关键词：微囊藻毒素，鸭胚，性腺，检测 Build of the Detection Method for Microcystins in the Gonad of Duck EmbryoLU Chen-Cen （Qiuzhen School of Huzhou Teachers College, Huzhou 313000） Abstract: Microcystins are a family of heptapeptides toxins with biological activity produced by freshwater cyanobacteria. Because of the fast development of modern industry, water pollution increasingly serious and eutrophication phenomenon growing heavily, the cyanobacteria proliferate and produce a large number of microcystins, it has seriously damaged animals and human healthy in the environment. In this experiment, we injected proper dose microcystins to the duck embryo, and used the ELISA test to detect the microcystins in the gonad of duck embryo at the different incubate date. The results showed that the microcystins could influence the duck growing development and accumulate in the gonad. The mortality of the duck embryos increased significantly when the injected dose was more than 0.1μg/egg. Key words: Microcystins, duck embryo, gonad, detect 目 录 1 引言…………………………………………………………………………………………1 2 材料与方法…………………………………………………………………………………2 2.1材料……………………………………………………………………………………2 2.1.1樱桃谷种鸭蛋 2.1.2微囊藻毒素 2.1.3 MC检测试剂盒 2.1.4化学药品和试剂 2.1.5主要仪器设备 2.1.6其他实验用具 2.1.7主要试剂的制备 2.1.8酶标仪的设置 2.1.9色谱柱的预处理 2.2方法和步骤……………………………………………………………………………4 2.2.1藻毒素的处理 2.2.2鸭胚的处理 2.2.3性腺组织中藻毒素的提取 2.2.4性腺组织中藻毒素的纯化 2.2.5性腺组织中藻毒素的检测 3 结果…………………………………………………………………………………………9 3.1鸭胚处理结果…………………………………………………………………………9 3.1.1种鸭蛋重量统计分析 3.1.2鸭胚孵化 3.1.3处理后鸭胚死亡情况 3.2性腺处理结果…………………………………………………………………………12 3.2.1性腺分离 3.2.2破碎、离心及纯化 3.3组织中微囊藻毒素的检测结果………………………………………………………13 4 讨论…………………………………………………………………………………………17 5 总结与展望…………………………………………………………………………………20 参考文献………………………………………………………