Development of Novel Toxin Detection Methodologies Applicable to Marine and Fresh Waters

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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. In this project, researchers at Lake Superior State University and the Texas A & M University at Corpus Christi will 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. There are four primary objectives: (1) to 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) to design and field validate a flexible MqPCR that can be tailored to specific HAB events; (3) to 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; and (4) to identify the role of poorly understood toxin-producing algae with U.S. freshwaters. This study is expected to 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.Broader Impacts. This work will further our understanding of the environmental drivers forcing shifts in HAB genera in the Laurentian Great Lakes and other important commercial, economical, and recreational freshwater resources systems across the USA. As different regions of the country have problems with different suites of toxins, the flexible MqPCR to be developed will give researchers and water managers the ability to tailor the MqPCR assay to suit specific bloom situations as needed. More generally, this work should significantly advance current understanding of the complex ecology of these events, enable better prediction of bloom toxicity, and improve the integration of HAB activities into existing state and federal monitoring and observational programs. The project would also provide for the training of a postdoctoral researcher.JOINT FUNDING BY NSF AND NIEHS: The original proposal on which this project is based (R01 ES021968-01) was submitted to the National Institutes of Environmental Health Sciences (NIH/NIEHS) in response to Funding Opportunity Announcement RFA-ES-11-013 , "Oceans, Great Lakes and Human Health (R01)", an opportunity jointly sponsored by NSF. This project is cooperatively funded through separate awards from NSF and NIEHS.