AQUATIC BIOMARKERS IN SITE CHARACTERIZATION AND REMEDIATION

该项目的主要目标是检测因接触水中单一或复杂混合物中的危险物质而造成的发育和生殖障碍。一旦检测到这些有害影响，将在一连串的毒物鉴定工作中对导致这种变化的样本进行分馏和处理。这些步骤将需要生物测定驱动的化学分析来确定复杂混合物的有毒成分(S)。尽管发育受损很重要，但使用这一点作为毒物鉴定评估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青竹，这项任务变得稍微容易了一些。青竹是一种小型硬骨鱼，长期以来一直被认为是发育生物学的模型。此外，新开发的纳米注射程序将用于精确地将胚胎暴露在有毒复杂混合物的候选成分中。一项协调的、重点关注的工作将与青竹基因组和cDNA文库以及分子工具包一起使用青竹组织、鲑鱼细胞系和组织，同时将专注于研究在体内被证明呈阳性的化合物的作用机制(S)。由于将被测试的一些化合物包括已被证明作为内分泌模拟物和/或内分泌干扰物的一些化合物，我们将包括中间性形成和间隔性成熟作为终点。协调的形态和生化/分子研究将解决对照和治疗个体性腺成熟所必需的蛋白质的特征。将对高速公路中途和外围使用的农用杀虫剂和除草剂进行测试。由于用于控制的化合物正在发生变化，我们将致力于分析新出现的化合物，拟除虫菊酯和酰基脲的代谢。在这一阶段的工作中，我们将重点创建一种前哨水生生物模型，用于检测常驻鱼类的暴露和有害影响。为了实现这一总体目标，提出了三个具体目标。1)利用先前校准的青竹成虫繁殖对以及配子和胚胎的暴露，确定危险废物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、幼虫和性成熟青竹的特定变化，显示暴露和影响的生物标记物；以及3)确定与化合物的生化和分子作用机制有关的新的生物标记物，包括那些被证明损害发育和/或生殖的生物标记物。

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.