Development of novel toxin detection methodologies applicable tomarine and fresh

开发适用于海洋和新鲜鱼类的新型毒素检测方法

• 批准号: 8554364
• 负责人: Judy A Westrick
• 金额: $ 6.6万
• 依托单位: LAKE SUPERIOR STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-24 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8554364
• 关键词: ANTX-a; Algae; Algal Blooms; Anabaena; Anabolism; Anatoxins; Biological Assay; Cells; Chemicals; Chemistry; Code; Complex; Country; Cyanobacterium; DNA Sequence; Data; Decision Modeling; Detection; Development; Discipline; Ecology; Ecosystem; Ensure; Environmental Risk Factor; Euglena; Event; Exposure to; Faculty; Frequencies; Fresh Water; Future; Gene Cluster; Gene Expression; Genetic; Genetic Markers; Genomics; Geographic state; Goals; Habitats; Health; Human; Knowledge; Lead; Mammals; Marines; Methodology; Methods; Microbiology; Modeling; Molecular; Monitor; Neurotoxins; Nutrient; Organism; Outcome; Polymerase Chain Reaction; Population; Postdoctoral Fellow; Process; Production; Reaction; Reporting; Research; Research Personnel; Resources; Rivers; Role; Sampling; Saxitoxin; Site; System; Techniques; Time; Toxic effect; Toxin; Water; anatoxin-a(s); cylindrospermopsin; data modeling; design; drinking; environmental change; exposed human population; flexibility; fluorophore; harmful algal blooms; improved; insight; member; microcystin; nodularin; novel; predictive modeling; programs; public health relevance; remediation; response; tool; toxic algae; toxic bloom; water quality; water treatment

DESCRIPTION (provided by applicant) Harmful algal blooms (HABs) can have severe ecological, economical, and human health impacts. At least 32 states have reported human or mammal exposures to HAB toxins. Furthermore, studies suggest that the frequency and intensity of HAB events have increased over the past few decades and will continue to increase this century. Knowledge gaps remain regarding the ecological drivers of these events which have hindered the development of successful mitigation strategies. Most HABs are comprised of several genera with each species containing toxic and non-toxic strains. This has made understanding the ecology of these events using traditional methods extremely difficult. Advances in molecular techniques (e.g. quantitative PCR) have facilitated the first steps towards a deeper understanding of these complex events although previous studies have generally focused on a single organism. The investigators' goal is to develop and employ a "flexible" real-time multiplex quantitative polymerase chain reaction (MqPCR) assay for the simultaneous quantification of co-occurring toxic HAB genera. This will facilitate the next important step in furthering our understanding of these complex events and will significantly improve the integration of HAB activities into existing state and federal monitoring and observational programs. The investigators have four primary objectives: 1) Develop MqPCR specific TaqMan primers and probes for the detection of anatoxin-a producing cyanobacteria as well as toxic eugleniods to incorporate into the existing MqPCR method. 2) Design and field validate a "flexible" MqPCR that can be tailored to specific HAB events. By designing both TaqMan primers and probes to be replicated in the same thermal cycling program, they will be able to select specific primer sets to match different bloom situations. 3) The investigators will use the MqPCR to investigate the response of microcystin/nodularin-, cylindrospermopsin-, saxitoxin-, and anatoxin-a-producing cyanobacteria as well as potentially toxic euglenoids to environmental variables individually, as well as in relation to each other, over the course of a bloom. They will sample from four sites across the nation each year that routinely develop dense blooms of mixed genera; western Lake Erie, Grand Lake St. Marys, OH, St. Johns River, FL, and Nueces River, TX. Samples will be collected weekly before, during and after bloom events (150 field samples minimum). The investigators will also collect nutrient data, physiochemical parameters and algal toxin data. 4) Identify the role of poorly understood toxin-producing algae with U.S. freshwaters. The three investigators (2 faculty members, one post-doctoral researcher) and a co-investigator's vast expertise in a number of disciplines (microbiology, genetics, and chemistry) will allow for a multi-disciplinary approach toward this research. The MqPCR technique developed will be a valuable tool for the rapid monitoring of HAB genera that can be applied across the marine- freshwater continuum. This study will provide important data for the modeling and prediction of future HAB events as well as the development of early warning networks to protect humans from exposure to HAB toxins. Public Health Relevance: Algal blooms cause serious water quality and human health issues; however, their ecology remains poorly understood. Using a cutting-edge genetic method the investigators will further their understanding of the environmental factors promoting these toxic algae. This will allow for the further development of successful predictive models and algal remediation strategies that will help protect humans from the negative human health effects of algal toxin exposure.

描述（由申请人提供） 有害藻华 (HAB) 会对生态、经济和人类健康产生严重影响。 至少有 32 个州报告了人类或哺乳动物接触 HAB 毒素的情况。此外，研究表明，HAB 事件的频率和强度在过去几十年中有所增加，并且本世纪将继续增加。 关于这些事件的生态驱动因素仍然存在知识差距，这阻碍了成功缓解策略的制定。大多数有害细菌由几个属组成，每个属都含有有毒和无毒菌株。这使得使用传统方法了解这些事件的生态变得极其困难。 尽管以前的研究通常集中在单一生物体上，但分子技术（例如定量 PCR）的进步促进了深入了解这些复杂事件的第一步。 研究人员的目标是开发和采用一种“灵活”的实时多重定量聚合酶链反应 (MqPCR) 测定法，用于同时定量共存的有毒 HAB 属。这将有助于我们进一步了解这些复杂事件，并显着改善 HAB 活动与现有活动的整合。 州和联邦监测和观察计划。 研究人员有四个主要目标：1) 开发 MqPCR 特异性 TaqMan 引物和探针，用于检测产生毒素的蓝细菌以及有毒眼虫，并将其纳入现有的 MqPCR 方法中。 2) 设计并现场验证可针对特定 HAB 事件定制的“灵活”MqPCR。 通过设计在同一热循环程序中复制的 TaqMan 引物和探针，他们将能够选择特定的引物组来匹配不同的水华情况。 3) 研究人员将使用 MqPCR 来研究在水华过程中，产生微囊藻毒素/球藻毒素、柱球藻毒素、石房蛤毒素和赤霉素 a 的蓝藻以及潜在有毒的眼虫对环境变量的反应，以及彼此之间的相互关系。 他们每年都会从全国各地的四个地点进行采样，这些地点通常会出现混合属的茂密花朵；伊利湖西部、俄亥俄州圣玛丽大湖、佛罗里达州圣约翰斯河和德克萨斯州纽西斯河。 将在开花事件之前、期间和之后每周收集样本（至少 150 个现场样本）。 研究人员还将收集营养数据、理化参数和藻类毒素数据。 4) 确定人们知之甚少的产毒藻类对美国淡水的影响。 三名研究人员（两名教职人员，一名博士后研究员）和一名联合研究人员在多个学科（微生物学、遗传学和化学）方面拥有丰富的专业知识，这使得这项研究能够采用多学科方法。开发的 MqPCR 技术将成为快速监测 HAB 属的宝贵工具，可应用于整个海洋-淡水连续体。这项研究将为未来 HAB 事件的建模和预测以及早期预警网络的开发提供重要数据，以保护人类免受 HAB 毒素的影响。 公共卫生相关性：藻华导致严重的水质和人类健康问题；然而，人们对它们的生态仍然知之甚少。研究人员将使用尖端的遗传方法进一步了解促进这些有毒藻类的环境因素。这将有助于进一步开发成功的预测模型和藻类修复策略，有助于保护人类免受藻类毒素暴露对人类健康的负面影响。

项目成果

Euglenophycin is produced in at least six species of euglenoid algae and six of seven strains of Euglena sanguinea.

• DOI: 10.1016/j.hal.2017.01.010
• 发表时间: 2017-03
• 期刊: Harmful algae
• 影响因子: 6.6
• 作者: Zimba PV;Huang IS;Gutierrez D;Shin W;Bennett MS;Triemer RE
• 通讯作者: Triemer RE

Augmenting anti-cancer natural products with a small molecule adjuvant.

用小分子佐剂增强天然产物的抗癌作用。

• DOI: 10.3390/md13010065
• 发表时间: 2014
• 期刊: Marine drugs
• 影响因子: 5.4
• 作者: Wahome,PaulG;Beauchesne,KevinR;Pedone,AnnaC;Cavanagh,John;Melander,Christian;Zimba,Paul;Moeller,PeterDR
• 通讯作者: Moeller,PeterDR

A Multiplex Analysis of Potentially Toxic Cyanobacteria in Lake Winnipeg during the 2013 Bloom Season.

对 2013 年水华季节温尼伯湖潜在有毒蓝藻的多重分析。

• DOI: 10.3390/toxins11100587
• 发表时间: 2019
• 期刊: Toxins
• 影响因子: 4.2
• 作者: McKindles,KatelynM;Zimba,PaulV;Chiu,AlexanderS;Watson,SusanB;Gutierrez,DanielleB;Westrick,Judy;Kling,Hedy;Davis,TimothyW
• 通讯作者: Davis,TimothyW

Phosphatidylcholine composition of pulmonary surfactant from terrestrial and marine diving mammals.

• DOI: 10.1016/j.resp.2015.02.004
• 发表时间: 2015-06
• 期刊: RESPIRATORY PHYSIOLOGY & NEUROBIOLOGY
• 影响因子: 2.3
• 作者: Gutierrez, Danielle B.;Fahlman, Andreas;Gardner, Manuela;Kleinhenz, Danielle;Piscitelli, Marina;Raverty, Stephen;Haulena, Martin;Zimba, Paul V.
• 通讯作者: Zimba, Paul V.

Benthic microalgae serve as the major food resource for porcelain crabs (Petrolisthes spp.) in oyster reefs: gut content and pigment evidence1.

底栖微藻是牡蛎礁瓷蟹（Petrolisthes spp.）的主要食物来源：肠道内容物和色素证据1。

• DOI: 10.1016/j.jembe.2016.06.005
• 发表时间: 2016
• 期刊: Journal of experimental marine biology and ecology
• 影响因子: 2
• 作者: Zimba,PaulV;Hill,ErinM;Withers,Kim
• 通讯作者: Withers,Kim