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

有害藻华（HABs）影响了许多沿海国家的大地理区域，威胁着环境和社会经济资源以及人类健康和福祉。该项目旨在利用2013年9月南加州聚鼓扁藻开花前、开花后和开花后的过滤样本，创建参考基因组和转录组数据库，并评估毒素产生和对人类细胞的影响。这些水华发生的时间难以预测，这使得目前的水华事件成为收集集中样本以评估这些偶发性事件对人类健康的潜在危害的机会。这些样本将用于DNA和RNA测序以及次级代谢物的提取，所产生的信息将被放置在参考库中，作为进一步检查存档样本的基础。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、转录组学和次级代谢组学参考库相结合，以进一步帮助解释数据。目标1描述已完成的收集和目前正在执行的收集。目的2从这些样品中提取、测序、组装和注释DNA和RNA。这些信息将与化学工作（Aim 3a）相结合，其中将调查样品是否产生次生代谢物（潜在毒素，包括亚致死）。

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.