【作者】 吴迪；

【导师】 单正军； 周立祥；

【作者基本信息】 南京农业大学 ， 环境工程， 2010， 硕士

【摘要】 环境内分泌干扰物对人类和生态环境的影响已经引起全球的广泛关注,特别是对于有机氯农药内分泌干扰的研究,国外已经进入深入阶段,而我国尚处于起步的阶段。本文选择水生生物斑马鱼为受试对象,参考OECD (Organization for Economic Cooperation and Development)内分泌干扰物筛选程序,对林丹和β-硫丹的内分泌干扰特性进行研究,以期为我国的环境内分泌干扰物的研究工作提供基础资料和有益借鉴。以斑马鱼为受试生物,研究在低浓度暴露条件下,林丹对斑马鱼体重体长、肝脏指数、性腺指数、产卵量、受精率、孵化率和卵黄蛋白原(Vtg)的影响。结果显示：经14d林丹(0.05μg/L、0.05μg/L、0.5μg/L)暴露后,处理组斑马鱼的体长体重与空白对照组无显著差异(P>0.05)；0.5μg/L处理组的雄性和雌性斑马鱼的肝脏指数均有显著增加(P<0.05),雄性斑马鱼的性腺指数有显著的降低(P<0.05),其余组无明显变化；林丹对斑马鱼产卵量和受精率表现出显著的抑制作用,浓度为0.05μg/L和0.5μg/L时,平均产卵量和受精率均显著的降低(P<0.05)；在林丹中暴露14d,各个浓度组的雄雌斑马鱼血浆中Vtg均没有显著的变化(P>0.05),SDS-PAGE电泳和Western-blotting结果表明斑马鱼Vtg的亚基为150kDa110kDa和86kDa,150KDa和110KDa为主要蛋白带。以斑马鱼为受试生物,研究β-硫丹对斑马鱼体重体长、肝脏指数、性腺指数、产卵量、受精率、孵化率和Vtg的影响。结果显示：设置浓度为0.01μg/L、0.05μg/L、0.2μg/L,在β-硫丹中暴露21天,各个浓度组的斑马鱼体长体重与对照组比较没有显著差异(P>0.05)；0.05μg/L、0.2μg/L两个处理浓度组雄性斑马鱼的肝脏指数均显著的增大(P<0.05),而雌鱼的肝脏指数与空白对照组比没有明显的变化(P>0.05)；雄性斑马鱼的性腺指数随着β-硫丹暴露浓度的增加有下降的趋势,0.2μg/L浓度暴露下,与空白对照组有显著差异(P<0.05),雌鱼性腺指数没有明显变化(P>0.05)；斑马鱼受精率随着β-硫丹暴露浓度的增加有下降的趋势,并且在0.2μg/L浓度时,与对照组有明显差异(P<0.05),各个暴露浓度组的孵化率都显著降低(P<0.05),表明了在β-硫丹中暴露21天,本实验设置的浓度对斑马鱼的受精率和孵化率都产生了显著的影响；各个暴露浓度组的雄性斑马鱼体内Vtg的含量与对照组比较是显著增加的,并且随着暴露浓度的增加而呈增大的趋势,表明p-硫丹可以诱导雄鱼体内产生Vtg,并且随着浓度的增加而增大。而雌鱼体内的Vtg含量并没有显著变化；SDS-PAGE和Western blotting结果表明斑马鱼能够发生特异性结合的Vtg亚基约为150kDa、110KDa和86kDa,150KDa(?)口110KDa为主要蛋白带。随着暴露浓度的增加,斑马鱼血浆内蛋白条带的宽度和深度都随着增加。本研究表明林丹和β-硫丹对斑马鱼表现出了内分泌干扰效应。更多还原

【Abstract】 The impacts of environmental endocrine disruptors on human and ecological environment have received worldwide attention in recent years. Compared with the advanced countries in this fidld of research specially to organochlorine, although China is still at an beginning stage. This study choosed zebrafish as the experimental animals. Refer to a short term screening test of OECD, we studied the endocrine disrupting effects of lindane and beta-endosulfan on zebrafish. This results could supply base datas and valuable experience to the research working of envirmonmental endocrine disruptors in our country.The effects of lindane on body length, body weight, hepaticosomatic index (HSI), gonadosomatic index (GSI), spawning and fertilization rates of zebrafish at low concentration were studied. The results showed that:after14-day exposure, the body length and body weight of zebrafish at all treated groups almost had no change when compared with the control groups. But the HSI of male and female zebrafish at0.5μg/L of lindane significantly increased (P<0.05) and the GSI of only male zebrafish were prominently reduced (P<0.05). Lindane remarkably inhibited zebrafish reproductive behavior and reduced the amount of average quantities of spawning and fertilization rate of zebrafish at0.05μg/L and0.5μg/L(P<0.05). The concentration of vitellogenin in the male and female zebrafish at all treated groups had no change afer14-day exposure (P>0.05).The results of SDS-PAGE and Western-blotting showed that the plasma of zebrafish contained three protein bands that reacted with mouse anti zebrafish vitellogenin monoclonal antibody.150KDa and100KDa were the major bands. Theses results demonstrated that lindane has a certain impact on the growth and reproduction of zebrafish at low concentration but no estrogenic impact on zebrafish.The effects of beta-endosulfan on body length, body weight, hepaticosomatic index (HSI), gonadosomatic index (GSI), hatching rates and fertilization rates of zebrafish at concentration of0.01μg/L,0.05μg/L,0.2μg/L were studied. The results showed that:the body length and body weight of male and female zebrafish at all treated groups had no significant change compared with the control groups after21-day exposure. The HSI of male zebrafish at0.05μg/L,0.2μg/L of beta-endosulfan significantly increased (P<0.05) but the female not.The GSI of only male zebrafish were prominently reduced at0.2μg/L (P <0.05).Beta-endosulfan inhibited zebrafish reproductive behavior and the hatching rates at0.2μg/L and fertilization rates at all concentrations had decreased remarkably (P<0.05) Although the concentration of vitellogenin in male zebrafish at all treated groups had increased significantly (P<0.05), the vitellogenin in female zebrafish were not affected by dietary concentrations used in this study.Three protein bands were proved to reacted with mouse anti zebrafish vitellogenin monoclonal antibody in the plasma of female and induced male zebrafish from the results of SDS-PAGE and Western-blotting. Theses results demonstrated that beta-endosulfan has reproductive toxicity and estrogenic impact on zebrafish.These results suggest that short-term exposure to lindane and beta-endosulfan have endocrine disrupting effects on zebrafish.更多还原