Using microbiomes as microsensors to forecast toxic algae blooms

项目摘要 有害赤潮正在成为沿海和内陆水质的最大威胁之一。 公共卫生。它们持续时间更长，发生频率更高，产生的有毒化学物质范围更广， 与过去几十年相比，对人类健康造成了负面影响，但我们无法预测藻类种群何时会 开花或产生毒素。由于回避是唯一的公共卫生策略，因此迫切需要制定 预警检测系统，在有害藻类爆发前(24小时)通知公众，以及2)识别 水柱中的化学或物理线索，预测最终生物标记物开发的HAB启动。 共生的海洋微生物群落(即微生物群)对光驱动的昼夜节律做出反应 当地光合作用藻类种群的变化和水化学变化。这导致了可检测到的移动 微生物组与藻类行为和化学物质相关的表达功能(即分子表型) 微扰。我们假设，依赖于时间的微生物组的分子表型对 有毒藻类水华之前的环境扰动，提供了对赤潮形成的早期检测。 这个项目的总体目标是追踪a)微生物群表达的多肽，以及b)水传播的 与华盛顿州伊斯特桑德水化学变化有关的代谢物，是唯一已知的具有 可以预见的是，哈布斯每年都会计时两次。目前还不能在空间或时间上预测其他赤潮 即使在1到2周的时间内也是如此。为了捕捉微生物组在开花前的分子特征， 我们将在第一年进行两次野外采集，时间定在开花开始之前。仅在下一年，我们就有 被授予24小时访问Eastound的私人码头的权限，实现了有史以来第一次开花前样本采集 为了了解赤潮的形成。我们已建立的高分辨率环境采样器将收集 微生物群和周围的水每4小时一次。由于细胞严格调节蛋白质的丰度，因此 与昼夜节律模式相关的蛋白质水平可以使用新的质量系统地量化 基于光谱分析的多肽分析。我们将评估&gt；20,000个多肽并测试其节律性和丢失情况 从而揭示了在HAB形成之前的关键时间点，并指示了局部 水或物理条件。利用获得的时间知识，我们将对其中的代谢物进行量化， 以及相邻的存档时间点，使用非靶向质谱学代谢组学，连接调制 多肽的节律性直接影响到微生物的代谢及其化学线索。这次会议的主要成果 拟议的工作将是鉴定在HAB形成和形成之前的肽和代谢物生物标记物。 对水华形成和毒素产生的化学控制有基本的了解。我们的长期目标 是开发一种快速的分子检测方法，可以检测到1)水的官能度 微生物组和/或2)有害藻华形成之前的关键水生代谢物，以便 防止人类接触水华毒素，从而保护人类健康。

Project Summary Harmful algal blooms (HABs) are becoming one of the greatest coastal and inland water quality threats to public health. They last longer, occur more frequently and produce a wider range of toxic chemicals that negatively impact human health than in past decades, yet we cannot forecast when an algal population will bloom or produce toxins. As avoidance is the only public health strategy, there is a critical need to 1) develop early warning detection systems to inform the public before (>24 hours) a harmful algae bloom, and 2) identify chemical or physical cues in the water column that forecast HAB initiation for eventual biomarker development. Co-occurring marine microbial communities (i.e., the microbiome) respond to light-driven circadian rhythms of local photosynthetic algal populations and changes in water chemistry. This results in a detectable shift in the microbiome’s expressed functions (i.e., molecular phenotype) that correlate to algal behavior and chemical perturbations. We hypothesize that the time-dependent, molecular phenotype of the microbiome responds to environmental perturbations that precede a toxic algal bloom, providing early detection of HAB formation. The overall objective for this project is to track a) peptides expressed by the microbiome, and b) waterborne metabolites associated with changing water chemistry in Eastsound, WA, the only known location that has predictably timed HABs twice every year. No other HABs can currently be spatially or temporally predicted even within a 1 to 2-week window. To capture the microbiome’s molecular signature of pre-bloom conditions, we will conduct two field collections in year 1 timed to precede bloom initiation. For the next year only, we have been granted 24hr access to a private dock in Eastsound, enabling the first-ever pre-bloom sample collection to understand HAB formation. Our established high-resolution environmental samplers will collect the microbiome and surrounding water every 4 hrs. Since cells tightly regulate protein abundance, changes in protein levels associated with circadian patterns can be systematically quantified using novel mass spectrometry-based peptide analyses. We will assess >20,000 peptides and test for rhythmicity and the loss thereof, revealing critical timepoints that precede HAB formation and indicate a significant change in the local water or physical conditions. Using the timing knowledge gained, we will then quantify metabolites in those, and adjacent, archived timepoints using untargeted mass spectrometric metabolomics, connecting modulations in peptide rhythmicity directly to microbiome metabolism and their chemical cues. The major outcome of the proposed work will be the identification of peptide and metabolite biomarkers that precede HAB formation and a fundamental understanding of chemical controls on bloom formation and toxin production. Our long-term goal is to develop a rapid molecular assay that detects the identified changes in 1) the functionality of the water microbiome and/or 2) crucial waterborne metabolites in advance of harmful algal bloom formation, in order to prevent human exposure to bloom toxins, thereby protecting human health.