Time-sensitive Collection and Analysis of a Marine-Derived Toxic Event

描述（由申请人提供）：对海洋来源的有毒事件进行时间敏感的收集和分析有害藻华（HAB）影响许多沿海国家的大片地理区域，并威胁环境和社会经济资源以及人类健康和福祉。该项目旨在使用来自当前（2013年9月）南加州多edrum的水华的水华前、水华和水华后过滤的样品来创建参考基因组和转录组数据库，并评估毒素产生和在人类细胞测定中的作用。这些水华爆发的时间不可预测，使得目前的水华事件成为一个收集浓缩样本的机会，以评估这些偶发事件对人类健康的潜在危害。样本将用于DNA和RNA测序以及次级代谢产物提取，创建的信息将被置于参考库中，作为存档样本进一步检查的基础。作为正在进行的收集的一部分，物理参数将被测量并与基因组学，转录组学和次级代谢组学参考库整合，以进一步帮助解释数据。目标1描述了已经收集的和正在进行的收集。目标2解决了从这些样品中提取、测序、组装和注释DNA和RNA。这一信息将与化学工作（目标3a）相结合，在化学工作中，将调查样品的次级代谢产物（潜在毒素，包括亚致死毒素）的产生情况。 使用质谱和核磁共振方法。将在4项表型体外试验中评价分离的次级代谢产物，用作其潜在细胞毒性、神经和免疫调节（即使在亚致死水平）的替代指标（目的3b）。总的来说，这项工作将通过创建一个参考图书馆来提供基础知识，这将有助于进一步检查自2008年以来收集的存档样本，其中包括水华和非水华样本。这些收集到的信息对于评估缓解战略和提供有毒事件的早期检测以限制对人类健康的影响至关重要，并将作为明年提交的R01赠款申请的基础。

DESCRIPTION (provided by applicant): Time-sensitive Collection and Analysis of a Marine-Derived Toxic Event Harmful algal blooms (HABs) impact many coastal states over large geographic areas and threaten environmental and socio-economical resources as well as human health and well-being. This project aims to use pre-bloom, bloom and post-bloom filtered samples from the current, September 2013, southern California bloom of Lingulodinium polyedrum to create a reference genome and transcriptome database, and assess toxin production and effect in human cell assays. The unpredictable timing of these blooms makes the current bloom event an opportunity to collect concentrated samples to evaluate potential harm to human health during these episodic events. The samples will be used for DNA and RNA sequencing and for secondary metabolite extractions, the information created will be placed in a reference library as the basis for further examination of archived samples. As part of ongoing collections, physical parameters will be measured and integrated with the genomic, transcriptomic and secondary metabolomic reference library to further aid in interpreting the data. Aim 1 describes the collections made and collections currently being performed. Aim 2 addresses the extraction, sequencing, assembly and annotation of DNA and RNA from these samples. This information will be coupled with a chemistry effort (Aim 3a) where samples will be investigated for their production of secondary metabolites (potential toxins, including sub-lethal) using mass spectrometry and NMR methods. The isolated secondary metabolites will be evaluated in four phenotypic in vitro assays used as a proxy for their potential cytotoxicity, neuro- and immuno-modulation even at sub-lethal levels (Aim 3b). In total, the work will provide fundamental knowledge by creating a reference library that will aid in further examination of archived samples collected since 2008, containing both bloom and non-bloom samples. This collected information is essential for evaluation of mitigation strategies and to provide early detection of toxic events in order to limit the impacts on human health, and will serve as the basis of an R01 grant application to be submitted next year.