Improving Biosecurity in Aquaculture using High Speed, Low cost, Lab on a Chip Micro-Cytometry for the Surveillance of Harmful Algal Blooms.

该项目将开发一种新的海水分析工具，以提高贝类渔业的生物安全性，因为贝类渔业容易受到与有害藻华有关的藻类生物毒素的污染。它将通过重新设计现有的芯片实验室（LOC）微细胞仪来实现这一目标，该微细胞仪可以对单个细胞在小于0.1mm宽的通道中以单个文件流动时进行计数和表征。它可以直接在海水样本中计数和区分特定的藻类细胞类型，包括英国本土的产生生物毒素的物种。该技术是现有细胞仪的低成本、小型化替代品，具有增强的功能和能力。它有潜力通过产生同步荧光和电阻抗测量来显著提高物种分辨率，后者在当前系统中是不存在的。基于这两个参数的目标物种的独特细胞图谱的生成提高了它们可以在含有具有相似形态特征的非目标物种的复杂样品中进行区分的准确性。该系统的其他优点包括没有鞘流（标准细胞仪中使用的清洁水来控制细胞在通道中的位置），这提高了吞吐量，消除了携带或产生清洁水的需要，并在必要时促进了流动通道的周期性解除阻塞（通过流动反转）。项目目标是由我们的“最终用户”之一和项目分包商环境、渔业和水产养殖科学中心（Cefas）共同构思和完善的，我们与该中心有着牢固而长期的合作关系。Cefas根据欧盟现行有关人类食用贝类质量的法规，进行法定有害藻华/藻类生物毒素监测。这需要定期监测水样中有害藻类的种类，并在必要时作为早期预警系统，刺激对贝类肉中藻类生物毒素的取样和分析。目前，送到中央实验室的水样是通过显微镜分析的。这一过程耗时且延误干预，导致人类健康风险增加，贝类产品损失给养殖业带来的潜在成本增加。因此，需要快速和近实时的分析系统来辅助/补充微观分析。为了解决这个问题，该项目将提供一种便携式台式LOC微细胞仪，用于快速计数和区分海水样品中的特定藻类细胞类型。该系统将能够由非专业人员使用自动数据分析软件操作，并将被我们的项目合作伙伴离岸贝类有限公司和环境局用作现场预警工具，以及我们的最终用户和分包商Cefas，以补充现有的水样分析（使用显微镜），以减少分析时间。项目产出还将补充现有的利用卫星成像的藻类监测方案，提供不受云层影响的现场数据。它将通过促进及时干预、直接带来经济收益和减少公共卫生风险而产生影响。该项目的产出将为未来开发一个强大的、可部署的系统奠定基础，该系统可以在包括海上在内的现场操作。这将提供近实时的贝类养殖水域分析，避免了从高风险地点收集样本的需要。关键词：有害藻华；micro-cytometer;芯片实验室（LOC）；微电脑系统;生物安全;贝类渔业;水产养殖

This project will develop a new seawater analysis tool to improve the biosecurity of shellfisheries, which are vulnerable to contamination with algal bio-toxins (poisons) that are associated with harmful algal blooms. It will do this by re-engineering an existing Lab on Chip (LOC) micro-cytometer that counts and characterises single cells as they flow in single file in a channel less than 0.1mm wide. It can count and discriminate specific algal cell types, including bio-toxin producing species native to the UK, directly in seawater samples. This technology represents a low cost, miniaturised alternative to existing cytometers with enhanced functionality and capability. It has the potential to significantly improve species resolution by generating synchronous fluorescence and electrical impedance measurements, the latter being absent from current systems. The generation of unique cytometric profiles for target species based on both parameters increases the accuracy with which they can be discriminated within complex samples containing non-target species with similar morphological characteristics. Other advantages of the system include the absence of a sheath flow (clean water used in standard cytometers to control cell position in the channel), which improves throughput, eliminates the need to carry or generate clean water and facilitates, where necessary, the periodic unblocking of the flow channel (by flow inversion). The project objectives were conceived with, and refined by one of our "end-users" and a project sub-contractor, the Centre for Environment, Fisheries and Aquaculture Sciences (Cefas), with whom we have a strong, long-standing collaborative relationship. Cefas carry out statutory harmful algal bloom / algal bio-toxin surveillance in line with current European Union regulations regarding the quality of shellfish produced for human consumption. This entails periodic monitoring of water samples for harmful algal species, and where necessary acts as an early warning system that stimulates the increased sampling and analysis of shellfish flesh for algal bio-toxins. Currently, water samples delivered to a centralised laboratory are analysed by microscopy. This process is time consuming and delays intervention, leading to an increased risk to human health and potential cost to the industry from loss of shellfish product. Accordingly, rapid and near-real time analysis systems are required to aid / complement microscopic analysis. To address this, the project will deliver a portable, bench-top LOC micro-cytometer for the rapid counting and discrimination of specific algal cell types in seawater samples. The system will be capable of being operated by non-specialist personnel using automated data analysis software and will be used by our project partners, Offshore Shellfish Ltd and the Environment Agency, as an in situ early warning tool, and our end-user and sub-contractor, Cefas, to complement existing water sample analysis (using microscopy), with a view to reducing analysis time. The project outputs will also complement existing algal surveillance programmes using satellite imaging by providing in situ data, uninterrupted by cloud cover. It will deliver impact by facilitating timely intervention, directly leading to economic gain and reduction in public health risks. The outputs from the project will underpin the future development of a robust, deployable system that can be operated in situ including offshore. This will provide near real-time analysis of shellfisheries waters, obviating the need to collect samples from high risk locations. Keywords: Harmful Algal Bloom (HAB); micro-cytometer; Lab on a Chip (LOC); Micro-systems; Biosecurity; Shellfisheries; Aquaculture