AQUATIC BIOMARKERS IN SITE CHARACTERIZATION AND REMEDIATION

这个项目的主要目标是探测由于接触水中的有害物质而造成的发育和繁殖受损，不论是单独接触还是作为复杂混合物接触。一旦检测到这些有害影响，将在一系列毒物鉴定练习中对导致这种变化的样品进行分离和处理。这些步骤将需要生物测定驱动的化学分析，以确定复杂混合物的有毒成分。尽管发育受损很重要，但将其作为毒物鉴定评价的终点将需要相当大的努力。然而，由于使用了一种独特的脊椎动物模型——medaka，一种小型硬骨鱼，长期以来被认为是发育生物学的模型，这项任务变得更容易了。此外，新开发的纳米注射程序将用于将胚胎精确暴露于有毒复杂混合物的候选成分中。利用medaka组织、鳟鱼细胞系和组织，利用medaka基因组和cDNA文库和分子工具包进行协调、集中的工作，同时集中研究在体内证明阳性的化合物的作用机制。由于将被测试的一些化合物包括已被证明具有内分泌模拟物和/或内分泌干扰物的作用，我们将把双性人的形成和性成熟的间隔作为终点。协调形态学和生化/分子研究将解决在对照和治疗个体中性腺成熟所需的蛋白质特征。将对高速公路中间和外围使用的农业农药和除草剂进行检测。由于用于防治的化合物在不断变化，我们将重点分析新出现的化合物，拟除虫菊酯类和酰基脲类的代谢。在这一阶段的工作中，我们将重点建立一个用于检测暴露和有害影响的常驻鱼类的哨兵水生生物模型。为实现总体目标，提出了三个具体目标。1)利用先前校准的medaka成年繁殖对和配子和胚胎暴露，确定危险废物复杂混合物中特定化合物的发育毒性和生殖影响；2)通过分子成像研究，确定胚胎、幼虫、幼体和性成熟medaka中表现出暴露和影响的生物标志物的特定变化；3)识别与化合物作用的生化和分子机制相关的新生物标志物，包括那些显示损害发育和/或生殖的化合物。

The major goal of this project is to detect impaired development and reproduction arising from exposure to hazardous substances, singly and as complex mixtures, in water. Once these deleterious effects are detected, fractionation and treatment of samples leading to such change will be performed in a series of toxicant identification exercises. These steps will require bioassay driven chemical analysis to identify the toxic component(s) of complex mixtures. Despite the importance of impaired development, use of this as an endpoint for toxicant identification evaluations will require considerable effort. However, this task is made somewhat easier by the use of a unique vertebrate model, the medaka, a small teleost fish long recognized as a model for developmental biology. In addition, newly developed nanoinjection procedures will be used for precise exposure of embryos to candidate components of toxic complex mixtures. A coordinated, focused effort with a medaka genomic and cDNA library and molecular toolkit using medaka tissues, trout cell lines and tissues will simultaneously concentrate on mechanism(s) of action of compounds proven positive in vivo. Since some of the compounds which will be tested include have been shown to act as endocrine mimics and/or endocrine disruptors, we shall include formation of intersex and interval to sexual maturity as an endpoint. Coordinated morphological and biochemical/molecular studies will address features of proteins necessary for gonadal maturation in control and treated individuals. Agricultural pesticides and herbicides used on highway medians and peripheries will be tested. Because the compounds used for control are changing, we shall devoted effort to analysis of metabolism of newly-emerging compounds, the pyrethroids and acyl ureas. In this phase of the work, we shall focus on creation of a sentinal aquatic organism model useful in detecting exposure and deleterious effect in resident fishes. To achieve the overall goal, three specific aims are proposed. 1) Determine developmental toxicity and reproductive effects of specific compounds in complex mixtures of hazardous wastes using previously calibrated medaka adult breeding pairs and exposure of gametes and embryos; 2) Define, via molecular imaging studies, specific alterations in embryos, larvae, juveniles and sexually mature medaka exhibiting biomarkers of exposure and effect; and, 3) Identify new biomarkers associated with the biochemical and molecular mechanisms of action of compounds including those shown to impair development and/or reproduction.