AQUATIC BIOMARKERS IN SITE CHARACTERIZATION AND REMEDIATION

该项目的主要目标是检测因接触水中单独和复杂混合物的危险物质而导致的发育和生殖受损。一旦检测到这些有害影响，将在一系列有毒物质鉴定工作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。尽管发育受损的重要性，但将其用作毒物鉴定评价的终点将需要相当大的努力。然而，这一任务是通过使用一个独特的脊椎动物模型，青鳉，一个小硬骨鱼长期以来被认为是发育生物学的模型变得更容易。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。使用青鳉组织、鳟鱼细胞系和组织的青鳉基因组和cDNA文库以及分子工具包的协调、集中的努力将同时集中于体内证明为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作为内分泌模拟物和/或内分泌干扰物，我们将包括中间性的形成和性成熟的间隔作为终点。协调的形态学和生物化学/分子研究将解决控制和治疗个体性腺成熟所需的蛋白质的特征。将对公路中间和周边使用的农药和除草剂进行测试。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，我们将集中精力建立一个用于检测常驻鱼类暴露和有害影响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)（2）通过分子成像研究，确定胚胎、幼体、幼体和性成熟青鳉中显示出接触和影响生物标志物的具体变化;和3）鉴定与化合物的生物化学和分子作用机制相关的新的生物标志物，包括显示损害发育和/或生殖的那些。

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.