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An On-Farm Workflow for Predictive Management of Paralytic Shellfish Toxin-Producing Harmful Algal Blooms for the Aquaculture Industry
Environmental Science & Technology ( IF 10.8 ) Pub Date : 2024-04-12 , DOI: 10.1021/acs.est.3c10502
Rendy Ruvindy 1 , Penelope A Ajani 1 , Sereena Ashlin 2 , Gustaaf Hallegraeff 3 , Kerstin Klemm 4 , Christopher J Bolch 3 , Sarah Ugalde 3, 5 , Mark Van Asten 6, 7 , Stephen Woodcock 1 , Matthew Tesoriero 1 , Shauna A Murray 1
Environmental Science & Technology ( IF 10.8 ) Pub Date : 2024-04-12 , DOI: 10.1021/acs.est.3c10502
Rendy Ruvindy 1 , Penelope A Ajani 1 , Sereena Ashlin 2 , Gustaaf Hallegraeff 3 , Kerstin Klemm 4 , Christopher J Bolch 3 , Sarah Ugalde 3, 5 , Mark Van Asten 6, 7 , Stephen Woodcock 1 , Matthew Tesoriero 1 , Shauna A Murray 1
Affiliation
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Paralytic shellfish toxins (PSTs) produced by marine dinoflagellates significantly impact shellfish industries worldwide. Early detection on-farm and with minimal training would allow additional time for management decisions to minimize economic losses. Here, we describe and test a standardized workflow based on the detection of sxtA4, an initial gene in the biosynthesis of PSTs. The workflow is simple and inexpensive and does not require a specialized laboratory. It consists of (1) water collection and filtration using a custom gravity sampler, (2) buffer selection for sample preservation and cell lysis for DNA, and (3) an assay based on a region of sxtA, DinoDtec lyophilized quantitative polymerase chain reaction (qPCR) assay. Water samples spiked with Alexandrium catenella showed a cell recovery of >90% when compared to light microscopy counts. The performance of the lysis method (90.3% efficient), Longmire’s buffer, and the DinoDtec qPCR assay (tested across a range of Alexandrium species (90.7–106.9% efficiency; r2 > 0.99)) was found to be specific, sensitive, and efficient. We tested the application of this workflow weekly from May 2016 to 30th October 2017 to compare the relationship between sxtA4 copies L–1 in seawater and PSTs in mussel tissue (Mytilus galloprovincialis) on-farm and spatially (across multiple sites), effectively demonstrating an ∼2 week early warning of two A. catenella HABs (r = 0.95). Our tool provides an early, accurate, and efficient method for the identification of PST risk in shellfish aquaculture.
中文翻译:
用于水产养殖业产生麻痹性贝类毒素的有害藻华的预测管理的农场工作流程
海洋甲藻产生的麻痹性贝类毒素(PST)对全球贝类产业产生了重大影响。在农场进行早期检测并进行最少的培训将为管理决策留出更多时间,从而最大限度地减少经济损失。在这里,我们描述并测试了基于sxtA4 (PST 生物合成中的初始基因)检测的标准化工作流程。工作流程简单且成本低廉,不需要专门的实验室。它包括 (1) 使用定制重力采样器进行水收集和过滤,(2) 用于样品保存和 DNA 细胞裂解的缓冲液选择,以及 (3) 基于sxtA区域的测定,DinoDtec 冻干定量聚合酶链式反应 ( qPCR)测定。与光学显微镜计数相比,添加链状亚历山大藻的水样显示细胞回收率 >90%。裂解方法(90.3% 效率)、Longmire 缓冲液和 DinoDtec qPCR 测定(在一系列亚历山大藻物种中进行测试(90.7–106.9% 效率; r 2 > 0.99))的性能被发现具有特异性、敏感性和稳定性。高效的。我们从 2016 年 5 月到 2017 年 10 月 30 日每周测试此工作流程的应用,以比较海水中的sxtA4拷贝 L –1与养殖场和空间上(跨多个地点)贻贝组织 ( Mytilus galloprovincialis )中的 PST 之间的关系,有效地证明了两个链状A. catenella HAB 的 2 周预警( r = 0.95)。我们的工具为识别贝类水产养殖中的 PST 风险提供了一种早期、准确和高效的方法。
更新日期:2024-04-12
中文翻译:
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用于水产养殖业产生麻痹性贝类毒素的有害藻华的预测管理的农场工作流程
海洋甲藻产生的麻痹性贝类毒素(PST)对全球贝类产业产生了重大影响。在农场进行早期检测并进行最少的培训将为管理决策留出更多时间,从而最大限度地减少经济损失。在这里,我们描述并测试了基于sxtA4 (PST 生物合成中的初始基因)检测的标准化工作流程。工作流程简单且成本低廉,不需要专门的实验室。它包括 (1) 使用定制重力采样器进行水收集和过滤,(2) 用于样品保存和 DNA 细胞裂解的缓冲液选择,以及 (3) 基于sxtA区域的测定,DinoDtec 冻干定量聚合酶链式反应 ( qPCR)测定。与光学显微镜计数相比,添加链状亚历山大藻的水样显示细胞回收率 >90%。裂解方法(90.3% 效率)、Longmire 缓冲液和 DinoDtec qPCR 测定(在一系列亚历山大藻物种中进行测试(90.7–106.9% 效率; r 2 > 0.99))的性能被发现具有特异性、敏感性和稳定性。高效的。我们从 2016 年 5 月到 2017 年 10 月 30 日每周测试此工作流程的应用,以比较海水中的sxtA4拷贝 L –1与养殖场和空间上(跨多个地点)贻贝组织 ( Mytilus galloprovincialis )中的 PST 之间的关系,有效地证明了两个链状A. catenella HAB 的 2 周预警( r = 0.95)。我们的工具为识别贝类水产养殖中的 PST 风险提供了一种早期、准确和高效的方法。