Establishment of method for evaluation for effects by endocrine disruptors on fish reproduction

内分泌干扰物对鱼类繁殖影响评价方法的建立

基本信息

批准号：
10356006
负责人：
YAMAUCHI Kohei
金额：
$ 12.61万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

(1) A comprehensive analytical method of endocrine disruptors (I.e., alkylphenols, phthalic acid esters, bisphenol A, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, DDTs) in environmental samples was developed. The method employs Soxhlet-extracion with dichloromethane and two-step-silica-gel and activated-charcoal column chromatography followed by determination on capillary GC equipped with MSD or ECD.(2) The change of olfactory cues was observed histologically and examined the effects on homing behavior prior to spawning in steroid hormone treated-salmon species.(3) High sensitive detection methods of estrogen receptor mRNA and vitellogenin mRNA of Japanese eel were established and these methods are useful tool for in vitro studies. While in order to establish a model system to study effects of estrogenic chemicals in the environment on aquatic animals, we examined the effects of a natural estrogen, 17 β-estradiol (E2) on early development of cyprinodont fish, mummichog.(4) Vitellogenins of male salmon, carp and flounder were mesured by sensitive ELISA. Mg/ml levels of vitellogenin were observed in male carp captured from seawage effluent and hundred μ g/ml levels were in male carp from the river of metropolitan.(5) The intersex condition which showed testes cotaining small numbers of ooytes, was observed among carp, flounder, red sea bream and mullet by histological anaylsis.(6) Effects of endocrine disrupting chemicals (bisphenol-A, nonylphenol and diphtalate) on reproductive ability of medaka were investigated. Adverse effects of estrogenic chemicals on reproductivity of medaka could be detected, so the materials and methods used were considered to be able to apply for a screening test method of mimic estrogen chemicals.

（1）建立了环境样品中内分泌干扰物（烷基酚、邻苯二甲酸酯、双酚A、多环芳烃、多氯联苯、滴滴涕）的综合分析方法。该方法采用二氯甲烷索氏萃取和两步法。硅胶和活性炭柱色谱法，然后在配备有毛细管气相色谱仪的毛细管气相色谱仪上测定MSD或ECD。(2)通过组织学观察嗅觉信号的变化，并研究其对类固醇激素处理的鲑鱼产卵前归巢行为的影响。(3)日本鳗鱼雌激素受体mRNA和卵黄蛋白原mRNA的高灵敏检测方法的建立，这些方法是体外研究的有用工具，同时为了建立一个模型系统来研究环境中雌激素化学物质对水生动物的影响，我们检查了天然雌激素的影响，17。 β-雌二醇(E2)对鲤鱼、mummichog早期发育的影响。(4)采用灵敏的ELISA法测定了雄性鲑鱼、鲤鱼和比目鱼的卵黄蛋白原，在从海水废水中捕获的雄性鲤鱼和百μl中观察到了Mg/ml的卵黄蛋白原水平。都市河雄性鲤鱼中的g/ml水平。(5)雌雄同体情况，睾丸含有少量通过组织学分析，在鲤鱼、比目鱼、真鲷和鲻鱼中观察到了卵母细胞。(6)研究了内分泌干扰物(双酚A、壬基酚和二邻苯二甲酸酯)对青鳉繁殖能力的影响。可以检测到青鳉，因此认为所用材料和方法能够适用于模拟雌激素化学品的筛选试验方法。