Development of a simple, low-cost device for sample collection and on-site preservation using a common oceanographic deployment platform

使用通用海洋学部署平台开发简单、低成本的样本采集和现场保存设备

• 批准号: 1924214
• 负责人: Peter Girguis
• 金额: $ 36.88万
• 依托单位: Harvard University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1924214&HistoricalAwards=false
• 关键词: Development; simple; low; cost; device

Over the past two decades, advanced genetic analysis techniques have accelerated society's understanding of marine microorganisms and the roles they play in ocean chemistry. This includes the discovery of new biological capacities and the recognition of their role in previously unknown chemical processes. Particularly, research on low-oxygen marine environments - including oxygen minimum zones, deep-sea hot springs, and other chemically-similar habitats - has revealed the critical influence they have on ocean health (for better and for worse). However, when sampling from such environments, researchers face biases that may arise due to biological changes or chemical degradation resulting from current means of sample collection and transport prior to analysis. This is a particular challenge for genetic materials that can degrade rapidly between sample collection and preservation. The technology development proposed here is aimed squarely at this challenge, with a secondary objective of providing the broader community with access to this technology. The development of a sampler that is compatible with a wide variety of water sampling platforms - at a very competitive price point - democratizes ocean science, bringing sampling tools to local, state, and federal coastal resource managers, non-profit organizations, teaching-focused institutions, and any investigator with a modest equipment budget. Further, this research will provide opportunities for engaging high school students in cutting edge research and engineering. While students are often exposed to microbiology and chemistry in the classroom, they are rarely afforded the opportunity to study the relationships among living organisms and their geochemical environment. The data from this research will contribute to student projects in an existing partnership between the researchers at Harvard University at the Cambridge Rindge and Latin School, a diverse public high school. Finally, in collaboration with the Harvard Museum of Natural History and CRLS, the researchers will conduct a Junior "XPRIZE" style competition. The participating students will build a water sampler to collect and preserve microbes at sea, and their devices will be tested alongside the researchers? prototype during the sea surface control experiments.The ocean is chemically and ecologically heterogeneous. For example, chemically reduced environments like those found at hydrothermal vents and hydrocarbon seeps are prominent features in the ocean. These locations host expansive microbial communities that play critical roles in processes from plankton health to planetary temperature. This is also true for the ocean's oxygen minimum zones (OMZs). There is a growing awareness of the biogeochemical connections that exist among the upper ocean and other well-oxygenated habitats, and those regions of the ocean that are depleted in or devoid of oxygen. Advancing the understanding of these relationships requires that researchers continue investigating A) the microbial diversity, distribution, and metabolic activity within these chemically-reducing environments; and B) the corresponding geochemical composition. To accomplish this, analysis of microbial gene expression products (e.g., RNA, proteins) and metabolites is essential. However, in sampling biomolecules from such environments, researchers are challenged to avoid biases that may arise due to changes in gene expression or chemical degradation during sample collection. This is especially challenging for molecules such as messenger RNA (mRNA) that can degrade rapidly between sample collection and preservation. This research aims to design, develop, and validate a low-cost, mRNA/protein/geochemical preservation system that will be "plug-n-play" with the Niskin rosettes that are found on most oceanographic research vessels. Broadly speaking, we propose an in situ sampler that can be mounted in lieu of a single Niskin bottle on a CTD rosette. Most importantly, it will be actuated via lanyard, like a standard Niskin bottle, thus eliminating the need for electrical interfacing. Once deployed to the appropriate depth and its lanyard is released, the system will begin its pre-programmed routine of collecting water samples and conducting the appropriate preservative steps. The device will also be capable of time-delayed sampling events, which will allow the user to re-position the CTD rosette to another depth and collect another sample. Deployment and operations testing will be conducted along a depth gradient at an off-shelf site in the major anoxic OMZ of the Eastern Tropical North Pacific. The sampler will be mounted onto a standard rosette and tested for successful firing, injection of preservative, and sample mixing at three depths spanning the anoxic nitrite maximum (~100-130 m), anoxic OMZ core (~300-700 m), and sub-OMZ (~1500 m). These three zones are known to support ecologically and taxonomically distinct microbial communities. In addition to system validation, effective analysis of microbial and chemical changes across this gradient will provide important information how microbes respond to changes in dissolved oxygen. Further, this will help to accurately delimit the boundaries of processes in OMZs and other redox-stratified habitats.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在过去的二十年中，先进的遗传分析技术加速了社会对海洋微生物及其在海洋化学中扮演的角色的理解。这包括发现新的生物学能力及其在以前未知的化学过程中的作用。特别是，对低氧海洋环境的研究 - 包括最小氧气区域，深海温泉和其他相似的栖息地 - 揭示了它们对海洋健康的关键影响（更加好转）。但是，当从这种环境中取样时，研究人员面临可能导致的偏见，这些偏见是由于当前的样品收集和运输方式引起的生物学变化或化学降解，然后进行分析。对于可以在样品收集和保存之间迅速降解的遗传材料，这是一个特别的挑战。此处提出的技术开发是针对这一挑战的，其次要目标是为更广泛的社区提供该技术的访问。与各种水样平台兼容的采样器的开发（在非常有竞争力的价格点）使海洋科学民主化，将采样工具带给地方，州和联邦沿海资源经理，非营利组织，注重教学的机构以及任何具有适度设备预算的研究人员。此外，这项研究将为高中生参与尖端研究和工程提供机会。尽管学生经常在课堂上接触微生物学和化学，但很少有机会研究生物体之间的关系及其地球化学环境。这项研究的数据将有助于哈佛大学研究人员在剑桥Rindge的研究人员与多元化公立高中拉丁语学校之间的现有合作伙伴关系。最后，与哈佛大学自然历史博物馆和CRL合作，研究人员将进行初级“ Xprize”风格竞赛。参与的学生将建造一个水采样器来收集和保存海上的微生物，他们的设备将与研究人员一起进行测试？海面控制实验期间的原型。海洋在化学和生态上是异质的。例如，化学减少的环境，例如在水热通风孔和碳氢化合物渗水处发现的环境是海洋中的突出特征。这些位置拥有广阔的微生物群落，这些群落在从浮游生活到行星温度的过程中起着关键作用。海洋的氧最小区（OMZ）也是如此。上海和其他氧化良好的栖息地之间存在的生物地球化学连接的认识越来越大，以及在氧气中耗尽或没有氧气的海洋地区。促进对这些关系的理解要求研究人员继续研究a）这些化学还原环境中的微生物多样性，分布和代谢活动； b）相应的地球化学组成。为此，必须分析微生物基因表达产物（例如RNA，蛋白质）和代谢产物。但是，在从这种环境中取样生物分子时，研究人员面临挑战，以避免由于样品收集过程中基因表达或化学降解而导致的偏见。对于可以在样品收集和保存之间迅速降解的分子（例如MESENGER RNA（mRNA）），这尤其具有挑战性。这项研究旨在设计，开发和验证低成本，mRNA/蛋白质/地球化学保存系统，该系统将与大多数海洋学研究容器中的Niskin Rosettes一起“插头”。从广义上讲，我们提出了一个原位采样器，该采样器可以安装，以代替CTD玫瑰花结上的一个Niskin瓶。最重要的是，它将像标准的Niskin瓶一样通过挂绳驱动，从而消除了对电气接口的需求。一旦部署到适当的深度及其挂绳被释放，该系统将开始其预编程的常规，以收集水样并进行适当的防腐剂。该设备还将能够进行时期的采样事件，这将使用户可以将CTD玫瑰花塞重新放置为另一个深度并收集另一个样本。部署和操作测试将沿深度梯度在东部热带北太平洋的主要缺氧OMZ的一个外部梯度沿深度梯度进行。采样器将安装在标准的玫瑰花结上，并在三个深度的最大氧化剂（〜100-130 m），缺氧OMZ核心（〜300-700 m）和sub-omz（〜1500 m）的三个深度和样品混合下进行测试。众所周知，这三个区域在生态和分类学上不同的微生物社区支持。除了系统验证外，对该梯度的微生物和化学变化的有效分析还将提供重要信息，微生物如何应对溶解氧的变化。此外，这将有助于准确地界定OMZ和其他氧化还原分层的栖息地中的过程界限。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子和更广泛影响的评估评估标准来通过评估来获得支持的。