Molecular Monitoring and Rapid-Response Characterization of Karenia brevis

短卡伦虫的分子监测和快速响应表征

• 批准号: 7682316
• 负责人: CHRISTOPHER D SINIGALLIANO
• 金额: $ 6.21万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7682316
• 关键词: Birds; Cell Count; Cell division; Cells; Cessation of life; Communities; Complex; Consensus; Consumption; Count; Data; Detection; Development; Environmental Impact; Environmental Risk Factor; Fishes; Florida; Flow Cytometry; Fluorescent in Situ Hybridization; Frequencies; Government; Harmful Algal Bloom; Health Professional; Hour; Human; Incidence; Laboratory culture; Light; Mammals; Marines; Measurement; Metabolic; Methodology; Methods; Mexico; Microscopic; Microscopy; Molecular; Monitor; Morphology; Phase; Photosynthesis; Phytoplankton; Polymerase Chain Reaction; Population; Process; Public Health; Rate; Research; Research Personnel; Respiration; Sampling; Seafood; Staging; Techniques; Testing; Time; Toxin; United States; aerosolized; base; coastal water; environmental health economics; gulf coast; harmful algal blooms; improved; indexing; killings; programs; respiratory; response; time use

The annual occurrence of Karenia brevis in the Gulk of Mexico is one of the most significant harmful algal blooms (HABs) in the coastal waters of the United States. This HAB species has a large economic, health, and environmental impact on Gulf coast communities, especially in Florida, through massive fish kills, deaths of marine mammals and birds, human consumption of toxin-contaminated seafood, and respiratory effects from aerosolized toxins. Over the past few decades the incidence of K. brevis blooms appears to be increasing in both frequency and duration. Current government-mandated programs for monitoring this HAB species must rely on microscopic cell counts. These counts are time consuming, have low sample through-put, and require highly-skilled microscopists who are experts at identifying and enumerating Karenia brevis cells of varying morphologies in complex natural phytoplankton assemblages. There is consensus among HAB researchers and public health professionals that monitoring efforts for HABs must evolve towards the use of rapid, accurate, sensitive, and objective molecular-based detection and quantitation methods. The proposed project will first utilize a combination of molecular techniques, such as Fluorescent In Situ Hybridization (FISH) and real-time quantitative polymerase chain reaction (qPCR), with traditional methods (light and fluorescent microscopy) to enumerate Karenia brevis cells in laboratory cultures. The objective of this initial phase is to test the efficacy of, and where feasible optimize, molecular methodologies with a short (3-5 hour) processing time for use in natural populations. Second, we will apply a combination of metabolic measurements (cell division, photosynthesis and respiration rates) and flow cytometry methodologies in cultures, and then natural popualtiosn, to improve our understanding of how environmental factors influence the metabolic state of K. brevis. Third, we will attempt to relate these indices to environmental parameters. The ultimate objective of this research is to facilitate the idenitfication and enumeration of this HAB species, and develop near real-time indices to assess the metabolic state of Karenia brevis in various stages of bloom development. Significance: It is anticipated that the results of this project will enhance our understanding of what factors contribute to Karenia bloom development. It will also synergistically enhance other collaborative HAB research programs through a rapid-response sampling program and sharing of samples and data.

每年在墨西哥湾出现的短卡伦虫是最严重的有害昆虫之一 美国沿海水域的藻华（HAB）。该HAB物种具有巨大的经济、 大量鱼类对墨西哥湾沿岸社区（尤其是佛罗里达州）的健康和环境影响 海洋哺乳动物和鸟类的捕杀、死亡、人类食用受毒素污染的海鲜，以及 雾化毒素对呼吸道的影响。在过去的几十年里，K. brevis 的发病率不断增加 频率和持续时间似乎都在增加。目前政府授权的计划 监测这种 HAB 物种必须依靠显微镜细胞计数。这些计数非常耗时，有 样品通量低，需要高技能的显微镜专家，他们是鉴定和鉴定方面的专家 列举复杂的天然浮游植物组合中不同形态的短卡伦藻细胞。 HAB 研究人员和公共卫生专业人员达成共识，即监测工作 HAB 必须朝着使用快速、准确、灵敏和客观的分子检测方向发展 和定量方法。 拟议的项目将首先利用分子技术的组合，例如荧光原位技术 使用传统方法进行杂交 (FISH) 和实时定量聚合酶链式反应 (qPCR) （光学和荧光显微镜）计数实验室培养物中的短卡伦尼亚细胞。的目标 这个初始阶段是测试分子方法的有效性，并在可行的情况下进行优化 用于自然群体的处理时间短（3-5小时）。其次，我们将组合应用 代谢测量（细胞分裂、光合作用和呼吸速率）和流式细胞术 文化中的方法论，然后是自然人口中的方法论，以提高我们对如何 环境因素影响短 K. brevis 的代谢状态。第三，我们将尝试将这些联系起来 环境参数指数。本研究的最终目的是促进识别 和计数该 HAB 物种，并开发近乎实时的指数来评估其代谢状态 短卡伦尼亚处于开花发育的各个阶段。 意义：预计该项目的结果将增强我们对以下内容的理解： 影响卡伦尼亚水华发育的因素。它还将协同增强其他协作 通过快速响应抽样计划以及样本和数据共享开展 HAB 研究计划。