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Amber Beecher Thesis.pdf (1.27 MB)

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Monitoring cyanotoxins in the western basin of Lake Erie using emerging technologies

Beecher, Amber A
http://rave.ohiolink.edu/etdc/view?acc_num=toledo1651835597744949

Abstract Details

2022, Master of Science, University of Toledo, Biology (Ecology).
The warm water temperatures, shallow depths and high nutrient inputs from surrounding farmlands make the Western Basin of Lake Erie a hotspot for Harmful Algal Blooms (HABs). HABs in Lake Erie are dominated by Microcystis sp. a type of cyanobacteria. Microcystis sp. frequently produce microcystin (MC), a hepatotoxin that affects the liver and other organs. MC concentrations can change rapidly in Lake Erie and drivers of toxin production are not well understood. The toxin concentration in water samples can be quantified in two well-established ways, including Enzyme Linked Immunosorbent Assay (ELISA) and mass spectrometry. On the other hand, Quantitative Polymerase Chain Reaction (qPCR) does not give concentrations of the toxin rather it gives the number of genes present that are able to produce the toxins. Monitoring for toxin concentrations in Lake Erie are traditionally done by vessel-based research and sample collection. Routine water samples (i.e. grab samples) collections are performed weekly at most but sometimes bi-weekly or less because of logistics such as cost, personnel requirements and especially weather conditions. With a decrease in sampling periods, traditional grab samples could potentially miss a peak in MC toxin. Non-traditional sampling method options, including enlisting the help of citizen scientist and using emerging technology that does not require weekly vessel-based research, needs to be explored to increase toxin concentration sampling in Lake Erie. The research in this thesis investigates monitoring techniques for MC toxin production in Lake Erie. This study aims to determine if SPATT bags are useful for aiding in the collection of MC toxin production and if SPATT bags are useful for different time intervals. SPATT bags were deployed from 6/25/2020-10/5/2020, bags were deployed for one-week and two-weeks through the sampling season. Results from the SPATT bags from the WBLE during the summer of 2020 were dominated by MC-LR> MC-RR> MC-YR> MC-LA, which composed 96% of the congeners present in SPATT bags. One-week and two-week bags had differing amounts of toxin concentrations possibly caused by the bags being colonized by toxin degrading bacteria. SPATT bags are useful in determining the congeners present in Lake Erie. Another aim for this study was to quantify the amount of MC producing genes (via qPCR) that were present at two sites compared to MC concentrations (via ELISA) in grab samples. Samples were taken weekly from 6/25/2020-10/5/2020. Results from the analysis showed that more consistent values were detected at the western site (Buoy), when comparing qPCR and ELISA, compared to the eastern site (Crib), which had higher relative MC concentrations (via ELISA) than the number of MC genes (via qPCR) from Microcystis. ELISA and qPCR values are correlated well at Buoy but not at Crib. The final aim of this study was to determine the best method to preserve DNA in filtered water samples that were not directly frozen at -80℃, implementing this method for citizen scientists. Water samples collected from pre-bloom and bloom periods were taken on 7/14/2021 and 8/19/2021, respectively. Samples were filtered onto a Veraspor 1.2 m 25 mm (Pall) filters and were either placed in silica gel, in a buffer solution or on an FTA card, then the treatments were either stored at room temperature (23℃), incubated at 37℃ for 2 weeks, or frozen at -80℃. Samples were then extracted, and the amount of DNA yield was compared to the standard. Results from this study indicated that silica gel was the most reliable method for preservation under all treatments. Using this method to get citizen scientists involved could increase the sample area and volume of samples in the future.
Thomas Bridgeman (Advisor)
Environmental Science
SPATT, Microcystis, Cyanotoxins

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Citations

  • Beecher, A. A. (2022). Monitoring cyanotoxins in the western basin of Lake Erie using emerging technologies [Master's thesis, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1651835597744949

    APA Style (7th edition)

  • Beecher, Amber. Monitoring cyanotoxins in the western basin of Lake Erie using emerging technologies . 2022. University of Toledo, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=toledo1651835597744949.

    MLA Style (8th edition)

  • Beecher, Amber. "Monitoring cyanotoxins in the western basin of Lake Erie using emerging technologies ." Master's thesis, University of Toledo, 2022. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1651835597744949

    Chicago Manual of Style (17th edition)

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