The Laurentian Great Lake, Lake Erie is an invaluable global resource and its watershed is home to over 11 million people. The pressures placed on the lake because of this high population caused Lake Erie to experience numerous environmental problems, including seasonal hypoxia and harmful algal blooms. While these topics have been widely studied in Lake Erie for over 40 years a more nuanced understanding of the interaction between phytoplankton and nutrient is needed to properly address the problems continuing to face the lake. In this study we combine classical limnological and cell growth experiments with modern molecular biological techniques and microscopy to more completely describe the aquatic microbial ecology of the lake. We used an oxalate rinse technique to examine the surface absorbed P pool of the toxic cyanobacterium Microcystis aeruginosa grown under a range of P conditions, as well as the general Lake Erie plankton assemblage. Our results suggest that while Microcystis is plastic in its cellular P needs, the ratio of intracellular to extracellular P remains stable across growth conditions. We describe the effect of the phosphonate herbicide glyphosate on the Lake Erie phytoplankton community using laboratory cell growth studies, field microcosm experiments and PCR amplification of a gene implicated in the breakdown of this compound from the environment. Results from these experiments suggest that the presence of glyphosate can affect community structure in multiple ways and may explain areas of unexplained phytoplankton diversity in coastal areas of Lake Erie. We also show heterotrophic bacteria are likely critical to the breakdown of glyphosate and further illustrate that understanding the context of the larger microbial community is critical to understanding the ecology of the constituent members of the community. Finally, we investigate the activity of the phytoplankton community in winter months with a focus on diatoms abundant in Lake Erie under the ice. We show these diatoms are active and that the winter bloom is a likely source of carbon important to seasonal hypoxia formation. Together, these studies significantly enrich our understanding of how phytoplankton influence important ecological processes in Lake Erie.

Inflows affect water quality, food web dynamics, and even the incidence of harmful algal blooms. It may be that inflows can be manipulated to create refuge habitat for biota trying to escape poor environmental conditions, such as fish populations in lakes during times of toxic Prymnesium parvum blooms. Water availability sometimes can be an issue, especially in arid climates, which limits this approach to management. Utilizing source water from deeper depths to displace surface waters, however, might effectively mimic inflow events. I test this notion by conducting in-lake mesocosm experiments with natural plankton communities where I manipulate hydraulic flushing. Results show that P. parvum cell density is reduced by 69%, and ambient toxicity completely ameliorates during pre-bloom conditions in the lake. During conditions of bloom development, population density is reduced by 53%, toxicity by 57%, and bloom proportions are never reached. There is minimal effect of these inflows on total phytoplankton and zooplankton biomass, and little effect on water quality. Shifts toward more rapidly growing phytoplankton taxa are observed, as are enhanced copepod nauplii. In other words, while inflows using deep waters suppress P. parvum bloom initiation and development, they are benign to other aspects of the lower food web and environment. The results from using deep lake water to suppress harmful algal blooms indicate this may be a promising management approach and further studies are recommended to test whether this mitigating effect can translate to a large-scale in-lake treatment.

This work explores the relationship between nutrient loading and changes in water quality in a sub-tropical, above-ground, off-stream municipal water supply reservoir, the C.W. Bill Young Regional Reservoir. The three source waters for the reservoir have varied but high levels of nutrients such as phosphorus and nitrogen. In other reservoirs, these nutrients have been linked to deterioration of water quality and increased expense in water treatment. The need to minimize excess nutrients results led to the primary research question: what allocation of withdrawals from the three sources will minimize the deterioration of water quality? To answer this question, the relationship between nutrient and other water quality data, such as temperature,phosphorus, chlorophyll a, Secchi depth, and trophic state indices were explored. Results indicate that temperature had a correlation with observed water quality. 27.9% of the variability in trophic state index as a function of Chlorophyll a was correlated with average temperature at one foot below water level. Correlation and regression models were developed using available time-series of linear and log-transformed water quality data to predict Chlorophyll a response. The parameters used in the model were selected from correlation matrices and from the P value in the multiple regression. The models developed were significant at P < 0.05. In the developed models, temperature was found to have greater predictive strength than nutrients indicating that this reservoir may be more strongly influenced by season and light than by nutrient limitation. Lastly, the US Army Corps of Engineers' eutrophication model, BATHTUB, was used to simulate different loading conditions and trophic response. The model results indicate that use of water the middle pool or lower pool of the Tampa Bypass Canal yield similar trophic states with the middle pool slightly lower. Use of water from the Alafia River yielded the highest trophic state and would be expected to have negative impacts on water quality.

This dissertation includes molecular and phytochemical investigations of the harmful, bloom-forming alga, Prymnesium parvum, including analysis of known polyketide metabolites as a function of salinity and growth. Initially, the development of molecular and phytochemical tools was necessary for the detection and quantification of P. parvum and its associated toxins. Suites of oligonucleotides and molecular beacons were designed for conventional and quantitative multiplex PCR to amplify four species- and gene-specific products simultaneously that were used for the detection and quantitation of P. parvum. This built-in redundancy provided increased confidence in reactions with the positive confirmation of four discrete products. Techniques were also developed for the chemical enrichment of toxins produced by P. parvum. Until now, isolation of “prymnesins” has never been reproduced. Polyketide prymnesins possess unique spectral properties that were used to generate an LC-MS fingerprint that comprised 13 ion species. Preliminary investigations using chemifluorimetric methods were also capable of detecting prymnesins in the pico- and nano-molar range. Environmental samples were tested as an independent assessment of these methods. Lastly, the roles of polyketide prymnesins were analyzed with respect to total hemolytic activity (HA) as a function of culture age and salinity. Variation in HA of supernatants was statistically significant relative to both variables (p << 0.05). Salinity was inversely related to HA wherein cultures growing in 5-25 psu were 150-200% more hemolytic. Total HA was inversely related to culture age during the first three weeks, but positively related to it during the next three weeks. Interestingly, no hemolysis was detected in fractions containing prymnesins from culture supernatants and the majority of hemolysins remained in the aqueous phase. Prymnesins extracted from cells varied significantly over the 6-week observation period (p << 0.05); HA was positively correlated during the first half and inversely related during the last half of the study. Salinity was directly related to HA from cell extracts, but these effects were not significantly different until the last three weeks. These investigations suggest that polyketide prymnesins are present at much lower quantities than previously believed, and they may not be the key compounds associated with hemolysis due to P. parvum. / text

Algae are important to marine and fresh-water ecosystems. However, some species of algae are harmful or even toxic. They can consume oxygen or block sunlight that is essential for other organisms to live. Indeed, some algae blooms can produce toxins that damage the health of the environment, plants, animals, and humans. Harmful algal blooms (HABs) which are often more green, brown, or dark-colored than red have spread along the coastlines and in the surface waters of the United States. Therefore, scientists are making great efforts to study HABs in order to maintain human and ecosystem health. Karenia brevis, the major harmful algal bloom dinoflagellate of the Gulf of Mexico, plays a destructive role in the region. Karenia brevis, responsible for Florida red tide, is the principle HAB dinoflagellate in the Gulf of Mexico. K. brevis blooms can produce brevetoxin: ladder-shaped polyether (LSP) compounds, which can lead to adverse human health effects, like reduced respiratory function through inhalation exposure, or neurotoxic shellfish poisoning through consumption of contaminated shellfish. The poisoning has been attributed to their affinity for voltage-sensitive sodium ion channels causing channel opening and depolarization of excitable cell membranes. Conservative estimate suggests that the economic impact from all harmful algal bloom events in the United States is at least $82 million/year. The public health costs occupy $37 million alone. The study presented herein utilized fluorescent and photolabile brevetoxin probes to demonstrate that brevetoxin localizes in the chloroplast of K. brevis where it binds to light harvest complex II (LHC II) and thioredoxin (Trx). It had been discovered that the TrxR/Trx system was inhibited by brevetoxin-2 (PbTx-2) with an IC50 of 25 µM. The mechanism of the inhibition was discussed in this work. The research also revealed that the K. brevis high-toxic and low-toxic strains have a significant difference in their ability, not only to produce brevetoxin, but also to perform NPQ and in the production of ROS. I compared and contrasted various metabolic and biochemical parameters in two strains of K. brevis which had a ten-fold difference in toxin content. The work could shed light on the physiological role that brevetoxin fills for K. brevis and may contribute to understanding the effect of ladder-shaped polyether compounds on both marine animals and exposed humans and shall inform improved treatments for brevetoxicosis.

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This study focused on Atmospheric Deposition (AD) loading on Utah Lake. Utah Lake is susceptible to Harmful Algal Blooms (HABs) because of its large surface area to volume ratio, proximity to Great Basin dust sources, and various wind patterns from close mountain ranges that blow AD towards the lake. In this study, we continued the collection and analysis of AD samples that started in 2017 and 2018, while reporting additional 2019 and 2020 data. We constructed a sampler on Utah Lake itself, which allowed us to better estimate how AD loads were distributed over the lake. An interpolation assumption was made in the previous studies that the amount of AD decreases exponentially as it passes onto the lake from the shore. Results from 5 months of Bird Island AD sampling on Utah Lake indicate that this assumption was incorrect. We performed statistical comparison tests on 2 variables: (1) the difference in AD between 2 table heights at the same site and (2) the difference in AD between a filtered sample and an unfiltered sample. We were able to statistically conclude that there was no difference in AD between 1-meter and 2-meter tall sample tables and that filtered AD samples had as much as 3 times lower concentration than unfiltered AD samples. In 2017, the total AD loading was estimated to be, on the high end, approximately 350 tons of total phosphorous (TP) and 460 tons of dissolved inorganic nitrogen (DIN) (Olsen JM, 2018). After making some changes to the interpolation methods, Joshua Reidhead in 2018 estimated AD loads of 153 tons of TP and 505 tons of DIN (Reidhead, 2019). With no changes to the 2018 sampling methods, but using an updated interpolation method, we determined the AD results for Utah Lake in 2019 to be 262 tons of TP and 1052 tons of DIN. After adjustments to the sampling tables, the bucket filters, and incorporating the Bird Island sampler results, we calculated the 2020 AD loading totals to be 133 tons of TP and 482 tons of DIN on the lake.

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