【作者】 那娜；

【导师】 张士璀；

【作者基本信息】 中国海洋大学 ， 海洋生物学， 2008， 博士

【摘要】 唑螨酯是日本在二十世纪九十年年代开发的一种新型吡唑类杀螨剂,由于其具有高选择性杀螨作用,在世界各国农业生产中的使用越来越广泛。对唑螨酯的代谢和残留研究表明唑螨酯在土壤和水体中具有较长的残留期,而且与其作用机理相似的农药均显示对鱼类高毒,并且与人类帕金森综合征有密切关系。因此唑螨酯对水体的污染以及对水生生物的毒性和人类健康的影响也越来越受到关注。本文采用了牙鲆活体及其鳃细胞系FG-9307作为体内外检测体系,系统研究了唑螨酯对于牙鲆这种海洋经济鱼类代表物种的急性毒性作用,并初步探讨了牙鲆及其鳃细胞系这两种体内外检测体系对污染物毒性检测的相关性。本研究首先探讨了唑螨酯对FG细胞系的体外急性毒性作用。研究发现:(1)三种检测方法测定的唑螨酯对FG细胞的48h IC50介于0.48-0.51μmol/L之间,96h IC50介于0.89-1.25μmol/L之间,说明唑螨酯对于体外培养鱼类细胞系具有毒性作用;同时也说明牙鲆鳃细胞系对于唑螨酯毒性作用比较敏感;(2)唑螨酯在0.1-30μmol/L浓度范围内对FG细胞生长具有明显抑制作用,并引起细胞形态明显改变;(3)唑螨酯影响了细胞抗氧化系统活性以及超氧阴离子的含量。经唑螨酯处理后的FG细胞,超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPX)和过氧化物酶(CAT)的活性均受到显著抑制,而超氧阴离子的含量明显升高。其次,本研究通过对牙鲆幼体LC50测定、肝脏抗氧化酶系统活性检测以及组织化学染色等研究方法,探讨了唑螨酯对牙鲆体内急性毒性作用。研究发现:(1)唑螨酯对牙鲆幼体48h和96h LC50分别为28.77 nmol/ L和11.74 nmol/ L;(2)唑螨酯影响了牙鲆肝脏抗氧化系统活性。肝脏抗氧化酶SOD、CAT和GPX活性在唑螨酯作用下出现明显的升高,随后又降低至低于正常的水平;(3)唑螨酯还能够造成牙鲆鳃、肝脏等组织器官的急性组织结构损伤,影响组织器官正常功能。最后,本研究对本实验室近年采用FG细胞系及玫瑰无须靶和牙鲆作为体内外毒性检测体系开展的环境污染物毒性研究数据资料进行了汇总分析,初步探讨了FG细胞系体外毒性与体内毒性检测结果之间的相关性。分析结果表明:体外毒性检测体系对环境污染物毒性检测敏感度低于体内检测体系,目前研究资料尚不能对牙鲆体内外检测体系的相关性关系做出明确判断,需要更多数据资料支持。总之,本研究表明唑螨酯对于牙鲆显示出高毒性。牙鲆及其鳃细胞系在近年来被越来越多应用于环境污染物毒性检测中,在本实验室开展的研究中也显示出对于环境污染物具有较高的敏感性,说明牙鲆及其鳃细胞系能够很好地反映环境污染物对海洋鱼类的体内外毒性效应。这对牙鲆及鳃细胞系作为环境污染物对海洋鱼类污染检测的模式动物提供了很好的理论依据。更多还原

【Abstract】 Fenpyroximate is a kind of mitochondrial electron transport inhibitors and acaricide, which have been developed by Japan from 1990s’, and now has been used more and more widely all over the world because of its high selective acaricide effect. Studies on the toxic metabolism and remaining of fenpyroximate in the environment show that, fenpyroximate could remain in soil and water for a relatively long period, and all the pesticides which have the same toxic mechanism to fenpyroximate show high toxicity to fish, and are closely related to human Parkinson’s Disease. So more and more attention has been paid to the toxicity of fenpyroximate on water environment, aqueous organisms, and human health. In this study, Flounder Paralichthy olivaceus and its gill cell line FG -9307 were selected as in vitro and in vivo testing systems, acute toxicity of fenpyroximate to this economical marine fish species was investigated, and the correlation of toxicities between these two testing systems were also discussed .In vitro toxicity of fenpyroximate to FG cells was investigated at first. The results showed that: (1) 48h IC50 tested by three methods was 0.48-0.51μmol/L, and 96h IC50 was 0.89-1.25μmol/L, indicating that fenpyroximate was toxic to in vitro fish cells, or FGl cells were highly sensitive to fenpyroximate; (2) fenpyroximate showed obvious inhibition effect on the proliferation of FG cells in the concentration range from 0.1 to 30μmol/L, and can cause FG cells obvious morphological changes; (3) fenpyroximate affected the activities of antioxidant enzyme system and the concentration of superoxide anion in FG cells. FG cells exposed to fenpyroximate exhibited prominent inhibition on the enzyme activities of SOD, CAT and GPX, and obvious increase of the concentration of superoxide anion in FG cells.Second, the in vivo toxicity of fenpyroximate to flounder was investigated by assay of the antioxidant enzyme activities and histochemistry examination. The results showed that, (1) 48 h and 96 h LC50 of fenpyroximate to young flounder were 28.77 nmol/ L and 11.74 nmol/ L, respectively; (2) fenpyroximate influenced r the antioxidation enzyme activities of flounder liver. the activities of SOD, CAT and GPX in the liver of flounder exposed to fenpyroximate increased obviously at the first several exposure hours and then were inhibited to be a level lower than that of control; (3) fenpyroximate can cause obvious damages to the structures of gill and liver of exposed flounder.In the end, we also analyzed the results of the toxicities of environmental pollutants using FG cell line, rosy barb(Puntius conchonius) and flounder as in vitro and in vivo toxicity testing systems, which have been carried out in our lab and investigated the correlation between in vitro and in vivo bioassay systems. The results indicated that the in vivo toxic testing system is more sensitive than in vitro system, and more data are needed to make a conclusion on the correlation of in vitro (FG cells) and in vivo (flounder) toxicity testing system更多还原