An assessment of a biosensor system for the quantification of microcystins in freshwater cyanobacterial blooms.

Microcystin-producing cyanobacterial blooms are a global issue threatening drinking water supplies and recreation on lakes and beaches. Direct measurement of microcystins is the only way to ensure waters have concentrations below guideline concentrations; however, analyzing water for microcystins takes several hours to days to obtain data. We tested LightDeck Diagnostics' bead beater cell lysis and two versions of the quantification system designed to give microcystin concentrations within 20 min and compared it to the standard freeze-thaw cycle lysis method and ELISA quantification. The bead beater lyser was only 30 % effective at extracting microcystins compared to freeze-thaw. When considering freeze-thaw samples analyzed in 2021, there was good agreement between ELISA and LightDeck version 2 (n = 152; R2 = 0.868), but the LightDeck slightly underestimated microcystins (slope of 0.862). However, we found poor relationships between LightDeck version 2 and ELISA in 2022 (n = 49, slopes 0.60 to 1.6; R2 < 0.6) and LightDeck version 1 (slope = 1.77 but also a high number of less than quantifiable concentrations). After the quantification issues are resolved, combining the LightDeck system with an already-proven rapid lysis method (such as microwaving) will allow beach managers and water treatment operators to make quicker, well-informed decisions.

Metagenomic and Metatranscriptomic Insights into Population Diversity of Microcystis Blooms: Spatial and Temporal Dynamics of mcy Genotypes, Including a Partial Operon That Can Be Abundant and Expressed.

Cyanobacterial harmful algal blooms (cyanoHABs) degrade freshwater ecosystems globally. Microcystis aeruginosa often dominates cyanoHABs and produces microcystin (MC), a class of hepatotoxins that poses threats to human and animal health. Microcystin toxicity is influenced by distinct structural elements across a diversity of related molecules encoded by variant mcy operons. However, the composition and distribution of mcy operon variants in natural blooms remain poorly understood. Here, we characterized the variant composition of mcy genes in western Lake Erie Microcystis blooms from 2014 and 2018. Sampling was conducted across several spatial and temporal scales, including different bloom phases within 2014, extensive spatial coverage on the same day (2018), and frequent, autonomous sampling over a 2-week period (2018). Mapping of metagenomic and metatranscriptomic sequences to reference sequences revealed three Microcystis mcy genotypes: complete (all genes present [mcyA-J]), partial (truncated mcyA, complete mcyBC, and missing mcyD-J), and absent (no mcy genes). We also detected two different variants of mcyB that may influence the production of microcystin congeners. The relative abundance of these genotypes was correlated with pH and nitrate concentrations. Metatranscriptomic analysis revealed that partial operons were, at times, the most abundant genotype and expressed in situ, suggesting the potential biosynthesis of truncated products. Quantification of genetic divergence between genotypes suggests that the observed strains are the result of preexisting heterogeneity rather than de novo mutation during the sampling period. Overall, our results show that natural Microcystis populations contain several cooccurring mcy genotypes that dynamically shift in abundance spatiotemporally via strain succession and likely influence the observed diversity of the produced congeners. IMPORTANCE Cyanobacteria are responsible for producing microcystins (MCs), a class of potent and structurally diverse toxins, in freshwater systems around the world. While microcystins have been studied for over 50 years, the diversity of their chemical forms and how this variation is encoded at the genetic level remain poorly understood, especially within natural populations of cyanobacterial harmful algal blooms (cyanoHABs). Here, we leverage community DNA and RNA sequences to track shifts in mcy genes responsible for producing microcystin, uncovering the relative abundance, expression, and variation of these genes. We studied this phenomenon in western Lake Erie, which suffers annually from cyanoHAB events, with impacts on drinking water, recreation, tourism, and commercial fishing.

Quantification of microcystin production and biodegradation rates in the western basin of Lake Erie.

Cyanobacterial biomass forecasts currently cannot predict the concentrations of microcystin, one of the most ubiquitous cyanotoxins that threaten human and wildlife health globally. Mechanistic insights into how microcystin production and biodegradation by heterotrophic bacteria change spatially and throughout the bloom season can aid in toxin concentration forecasts. We quantified microcystin production and biodegradation during two growth seasons in two western Lake Erie sites with different physicochemical properties commonly plagued by summer Microcystis blooms. Microcystin production rates were greater with elevated nutrients than under ambient conditions and were highest nearshore during the initial phases of the bloom, and production rates were lower in later bloom phases. We examined biodegradation rates of the most common and toxic microcystin by adding extracellular stable isotope-labeled microcystin-LR (1 µg L-1), which remained stable in the abiotic treatment (without bacteria) with minimal adsorption onto sediment, but strongly decreased in all unaltered biotic treatments, suggesting biodegradation. Greatest biodegradation rates (highest of -8.76 d-1, equivalent to the removal of 99.98% in 18 h) were observed during peak bloom conditions, while lower rates were observed with lower cyanobacteria biomass. Cell-specific nitrogen incorporation from microcystin-LR by nanoscale imaging mass spectrometry showed that a small percentage of the heterotrophic bacterial community actively degraded microcystin-LR. Microcystin production and biodegradation rates, combined with the microcystin incorporation by single cells, suggest that microcystin predictive models could be improved by incorporating toxin production and biodegradation rates, which are influenced by cyanobacterial bloom stage (early vs. late bloom), nutrient availability, and bacterial community composition.

Effectiveness of a fixed-depth sensor deployed from a buoy to estimate water-column cyanobacterial biomass depends on wind speed.

Water quality sondes have the advantage of containing multiple sensors, extended deployment times, high temporal resolution, and telecommunication with stakeholder accessible data portals. However, sondes that are part of buoy deployments often suffer from typically being fixed at one depth. Because water treatment plants are interested in water quality at a depth of the water intake and other stakeholders (ex. boaters and swimmers) are interested in the surface, we examined whether a fixed depth of approximately 1 m could cause over- or under-estimation of cyanobacterial biomass. We sampled the vertical distribution of cyanobacteria adjacent to a water quality sonde buoy in the western basin of Lake Erie during the summers of 2015-2017. A comparison of buoy cyanobacteria RFU (Relative Fluorescence Unit) at 1 m to cyanobacteria chlorophyll a (chla) measured throughout the water column showed occurrences when the buoy both under and overestimated the cyanobacteria chla at specific depths. Largest differences between buoy measurements and at-depth grab samples occurred during low wind speeds (< 4.5 m/sec) because low winds allowed cyanobacteria to accumulate at the surface above the buoy's sonde. Higher wind speeds (> 4.5 m/sec) resulted in better agreement between the buoy and at-depth measurements. Averaging wind speeds 12 hr before sample collection decreased the difference between the buoy and at-depth samples for high wind speeds but not low speeds. We suggest that sondes should be placed at a depth of interest for the appropriate stakeholder group or deploy sondes with the ability to sample at various depths.

Record-setting algal bloom in Lake Erie caused by agricultural and meteorological trends consistent with expected future conditions.

In 2011, Lake Erie experienced the largest harmful algal bloom in its recorded history, with a peak intensity over three times greater than any previously observed bloom. Here we show that long-term trends in agricultural practices are consistent with increasing phosphorus loading to the western basin of the lake, and that these trends, coupled with meteorological conditions in spring 2011, produced record-breaking nutrient loads. An extended period of weak lake circulation then led to abnormally long residence times that incubated the bloom, and warm and quiescent conditions after bloom onset allowed algae to remain near the top of the water column and prevented flushing of nutrients from the system. We further find that all of these factors are consistent with expected future conditions. If a scientifically guided management plan to mitigate these impacts is not implemented, we can therefore expect this bloom to be a harbinger of future blooms in Lake Erie.

Quantifying and Reducing Uncertainty in Estimated Microcystin Concentrations from the ELISA Method.

We discuss the uncertainty associated with a commonly used method for measuring the concentration of microcystin, a group of toxins associated with cyanobacterial blooms. Such uncertainty is rarely reported and accounted for in important drinking water management decisions. Using monitoring data from Ohio Environmental Protection Agency and from City of Toledo, we document the sources of measurement uncertainty and recommend a Bayesian hierarchical modeling approach for reducing the measurement uncertainty. Our analysis suggests that (1) much of the uncertainty is a result of the highly uncertain "standard curve" developed during each test and (2) the uncertainty can be reduced by pooling raw test data from multiple tests. Based on these results, we suggest that estimation uncertainty can be effectively reduced through the effort of either (1) regional regulatory agencies by sharing and combining raw test data from regularly scheduled microcystin monitoring program or (2) the manufacturer of the testing kit by conducting additional tests as part of an effort to improve the testing kit.

Field and laboratory studies of fluorescence-based technologies for real-time tracking of cyanobacterial cell lysis and potential microcystins release.

Elevated levels of dissolved microcystins (MCs) in source water due to rapid cell lysis of harmful cyanobacterial blooms may pose serious challenges for drinking water treatment. Catastrophic cell lysis can result from outbreaks of naturally-occurring cyanophages - as documented in Lake Erie during the Toledo water crisis of 2014 and in 2019, or through the application of algaecides or water treatment chemicals. Real-time detection of cyanobacterial cell lysis in source water would provide a valuable tool for drinking water plant and reservoir managers. In this study we explored two real-time fluorescence-based devices, PhycoSens and PhycoLA, that can detect unbound phycocyanin (uPC) as a potential indication of cell lysis and MCs release. The PhycoSens was deployed at the Low Service pump station of the City of Toledo Lake Erie drinking water treatment plant from July 15 to October 19, 2022 during the annual cyanobacteria bloom season. It measured major algal groups and uPC in incoming lake water at 15-min intervals during cyanobacteria dominant and senescence periods. Intermittent uPC detections from the PhycoSens over a three-month period coincided with periods of increasing proportions of extracellular MCs relative to total (intracellular and extracellular) MCs, indicating potential for uPC use as an indicator of cyanobacterial cell integrity. Following exposures of laboratory-cultured MCs-producing Microcystis aeruginosa NIES-298 (120 µg chlorophyll/L) to cyanophage Ma-LMM01, copper sulfate (0.5 and 1 mg Cu/L), sodium carbonate peroxyhydrate (PAK® 27, 6.7 and 10 mg H2O2/L), and potassium permanganate (2.5 and 4 mg/L), appearance of uPC coincided with elevated fractions of extracellular MCs. The PhycoLA was used to monitor batch samples collected daily from Lake Erie water exposed to algaecides in the laboratory. Concurrence of uPC signal and surge of dissolved MCs was observed following 24-h exposures to copper sulfate and PAK 27. Overall results indicate the appearance of uPC is a useful indicator of the onset of cyanobacterial cell lysis and the release of MCs when MCs are present.

Declines in ice cover are accompanied by light limitation responses and community change in freshwater diatoms.

The rediscovery of diatom blooms embedded within and beneath the Lake Erie ice cover (2007-2012) ignited interest in psychrophilic adaptations and winter limnology. Subsequent studies determined the vital role ice plays in winter diatom ecophysiology as diatoms partition to the underside of ice, thereby fixing their location within the photic zone. Yet, climate change has led to widespread ice decline across the Great Lakes, with Lake Erie presenting a nearly "ice-free" state in several recent winters. It has been hypothesized that the resultant turbid, isothermal water column induces light limitation amongst winter diatoms and thus serves as a competitive disadvantage. To investigate this hypothesis, we conducted a physiochemical and metatranscriptomic survey that spanned spatial, temporal, and climatic gradients of the winter Lake Erie water column (2019-2020). Our results suggest that ice-free conditions decreased planktonic diatom bloom magnitude and altered diatom community composition. Diatoms increased their expression of various photosynthetic genes and iron transporters, which suggests that the diatoms are attempting to increase their quantity of photosystems and light-harvesting components (a well-defined indicator of light limitation). We identified two gene families which serve to increase diatom fitness in the turbid ice-free water column: proton-pumping rhodopsins (a potential second means of light-driven energy acquisition) and fasciclins (a means to "raft" together to increase buoyancy and co-locate to the surface to optimize light acquisition). With large-scale climatic changes already underway, our observations provide insight into how diatoms respond to the dynamic ice conditions of today and shed light on how they will fare in a climatically altered tomorrow.

Microcystin congeners in Lake Erie follow the seasonal pattern of nitrogen availability.

Cyanobacteria harmful algal blooms produce many toxic secondary metabolites called cyanotoxins. The most studied group of cyanotoxins are microcystins (MC), with over 300 congeners reported. MC-LR is the most studied congener because of its abundance and toxicity. Recent toxicology studies suggest that more hydrophobic MC congeners such as MC-LA, MC-LF, and MC-LW may be less abundant but up to seven times more toxic than MC-LR, whereas, MC-RR's toxicity is only one-fifth that of MC-LR. Hence, understanding the environmental stressors that change the MC congener profile is critical to assessing the negative impact on environmental and human health. A two-year field and experimental study investigated seasonal and spatial changes of MC congener profiles in the western basin of Lake Erie. Both studies showed that nitrogen enrichment favored the production of nitrogen-rich MC-RR (C49H75N13O12). The field study showed that nitrogen depletion favored the low-nitrogen MC-LA (C46H67N7O12). MC-LR (a medium N level, C49H75N10O12) accounted for ∼30% to 50% of the total MC concentration and was stable across nitrogen concentrations. Using the relative toxicity and concentrations of each MC congener, both LC-MS/MS and ELISA overestimated the toxicity early bloom (July) and underestimated it late bloom (September). On 24 July 2019, highly toxic MC-LW and MC-LF were detected at nearshore stations with relative toxicity exceeding drinking water standards. This study demonstrated that the less toxic, high nitrogen MC-RR dominated under nitrogen-replete conditions in the early season, whereas the more toxic, less nitrogen MC-LA dominated under nitrogen-limited conditions later in the season.

Metatranscriptomic Sequencing of Winter and Spring Planktonic Communities from Lake Erie, a Laurentian Great Lake.

Previous reports suggest planktonic and under-ice winter microbial communities in Lake Erie are dominated by diatoms. Here, we report the assembled metatranscriptomes of 79 Lake Erie surface water microbial communities spanning both the winter (28 samples) and spring (51 samples) months over spatial, temporal, and climatic gradients in 2019 through 2020.

Changes in Microbiome Activity and Sporadic Viral Infection Help Explain Observed Variability in Microcosm Studies.

The environmental conditions experienced by microbial communities are rarely fully simulated in the laboratory. Researchers use experimental containers ("bottles"), where natural samples can be manipulated and evaluated. However, container-based methods are subject to "bottle effects": changes that occur when enclosing the plankton community that are often times unexplained by standard measures like pigment and nutrient concentrations. We noted variability in a short-term, nutrient amendment experiment during a 2019 Lake Erie, Microcystis spp. bloom. We observed changes in heterotrophic bacteria activity (transcription) on a time-frame consistent with a response to experimental changes in nutrient availability, demonstrating how the often overlooked microbiome of cyanobacterial blooms can be altered. Samples processed at the time of collection (T0) contained abundant transcripts from Bacteroidetes, which reduced in abundance during incubation in all bottles, including controls. Significant biological variability in the expression of Microcystis-infecting phage was observed between replicates, with phosphate-amended treatments showing a 10-fold variation. The expression patterns of Microcystis-infecting phage were significantly correlated with ∼35% of Microcystis-specific functional genes and ∼45% of the cellular-metabolites measured across the entire microbial community, suggesting phage activity not only influenced Microcystis dynamics, but the biochemistry of the microbiome. Our observations demonstrate how natural heterogeneity among replicates can be harnessed to provide further insight on virus and host ecology.

Elevated pH Conditions Associated With Microcystis spp. Blooms Decrease Viability of the Cultured Diatom Fragilaria crotonensis and Natural Diatoms in Lake Erie.

Cyanobacterial Harmful Algal Blooms (CyanoHABs) commonly increase water column pH to alkaline levels ≥9.2, and to as high as 11. This elevated pH has been suggested to confer a competitive advantage to cyanobacteria such as Microcystis aeruginosa. Yet, there is limited information regarding the restrictive effects bloom-induced pH levels may impose on this cyanobacterium's competitors. Due to the pH-dependency of biosilicification processes, diatoms (which seasonally both precede and proceed Microcystis blooms in many fresh waters) may be unable to synthesize frustules at these pH levels. We assessed the effects of pH on the ecologically relevant diatom Fragilaria crotonensis in vitro, and on a Lake Erie diatom community in situ. In vitro assays revealed F. crotonensis monocultures exhibited lower growth rates and abundances when cultivated at a starting pH of 9.2 in comparison to pH 7.7. The suppressed growth trends in F. crotonensis were exacerbated when co-cultured with M. aeruginosa at pH conditions and cell densities that simulated a cyanobacteria bloom. Estimates demonstrated a significant decrease in silica (Si) deposition at alkaline pH in both in vitro F. crotonensis cultures and in situ Lake Erie diatom assemblages, after as little as 48 h of alkaline pH-exposure. These observations indicate elevated pH negatively affected growth rate and diatom silica deposition; in total providing a competitive disadvantage for diatoms. Our observations demonstrate pH likely plays a significant role in bloom succession, creating a potential to prolong summer Microcystis blooms and constrain diatom fall resurgence.

Roles of Nutrient Limitation on Western Lake Erie CyanoHAB Toxin Production.

Cyanobacterial harmful algal bloom (CyanoHAB) proliferation is a global problem impacting ecosystem and human health. Western Lake Erie (WLE) typically endures two highly toxic CyanoHABs during summer: a Microcystis spp. bloom in Maumee Bay that extends throughout the western basin, and a Planktothrix spp. bloom in Sandusky Bay. Recently, the USA and Canada agreed to a 40% phosphorus (P) load reduction to lessen the severity of the WLE blooms. To investigate phosphorus and nitrogen (N) limitation of biomass and toxin production in WLE CyanoHABs, we conducted in situ nutrient addition and 40% dilution microcosm bioassays in June and August 2019. During the June Sandusky Bay bloom, biomass production as well as hepatotoxic microcystin and neurotoxic anatoxin production were N and P co-limited with microcystin production becoming nutrient deplete under 40% dilution. During August, the Maumee Bay bloom produced microcystin under nutrient repletion with slight induced P limitation under 40% dilution, and the Sandusky Bay bloom produced anatoxin under N limitation in both dilution treatments. The results demonstrate the importance of nutrient limitation effects on microcystin and anatoxin production. To properly combat cyanotoxin and cyanobacterial biomass production in WLE, both N and P reduction efforts should be implemented in its watershed.

The Lake Erie HABs Grab: A binational collaboration to characterize the western basin cyanobacterial harmful algal blooms at an unprecedented high-resolution spatial scale.

Monitoring of cyanobacterial bloom biomass in large lakes at high resolution is made possible by remote sensing. However, monitoring cyanobacterial toxins is only feasible with grab samples, which, with only sporadic sampling, results in uncertainties in the spatial distribution of toxins. To address this issue, we conducted two intensive "HABs Grabs" of microcystin (MC)-producing Microcystis blooms in the western basin of Lake Erie. These were one-day sampling events during August of 2018 and 2019 in which 100 and 172 grab samples were collected, respectively, within a six-hour window covering up to 2,270 km2 and analyzed using consistent methods to estimate the total mass of MC. The samples were analyzed for 57 parameters, including toxins, nutrients, chlorophyll, and genomics. There were an estimated 11,513 kg and 30,691 kg of MCs in the western basin during the 2018 and 2019 HABs Grabs, respectively. The bloom boundary poses substantial issues for spatial assessments because MC concentration varied by nearly two orders of magnitude over very short distances. The MC to chlorophyll ratio (MC:chl) varied by a factor up to 5.3 throughout the basin, which creates challenges for using MC:chl to predict MC concentrations. Many of the biomass metrics strongly correlated (r > 0.70) with each other except chlorophyll fluorescence and phycocyanin concentration. While MC and chlorophyll correlated well with total phosphorus and nitrogen concentrations, MC:chl correlated with dissolved inorganic nitrogen. More frequent MC data collection can overcome these issues, and models need to account for the MC:chl spatial heterogeneity when forecasting MCs.

Detection and quantification of Microcystis spp. and microcystin-LR in Western Lake Erie during the summer of 2007.

Microcystis spp. blooms have occurred annually in western Lake Erie since about 1995. Microcystis produce a group of toxins known as microcystins which can be harmful to livestock and to humans. In this study, surface water samples were collected from six sites during six sampling events from July to October in 2007. In situ environmental data (e.g. pH, temperature) and laboratory analyses (e.g. nutrients) were carried out to characterize the six sites. The Microcystis spp. density ranged from 10(2) to 10(7) cells/ml. Microcystin-LR concentration of 20 of all 36 samples were below the detection limit (0.15-5 ppb), while the microcystin-LR concentration in the 16 remaining samples ranged from 0.5 to 3 x 10(3) microg per gram dry weight. The aim of this research was to investigate the relationships between sampling location, environmental parameters, Microcystis spp. concentration, and microcystin-LR concentration. The results suggest that temperature, nutrient concentration, turbidity, and wind speed and direction (P<0.05) are factors which affected Microcystis spp. density. Sampling site 8M, located 13 m from the Maumee River, provided an advantage for Microcystis spp. growth, presumably due to intermediate water depth (5.5 m) combined with impact from the river. No relationship was found between Microcystis spp. density and microcystin-LR concentration. Temperature, nutrient concentration and DO (P<0.05) were associated with the production of microcystin-LR.

Science meets policy: A framework for determining impairment designation criteria for large waterbodies affected by cyanobacterial harmful algal blooms.

Toxic cyanobacterial harmful algal blooms (cyanoHABs) are one of the most significant threats to the security of Earth's surface freshwaters. In the United States, the Federal Water Pollution Control Act of 1972 (i.e., the Clean Water Act) requires that states report any waterbody that fails to meet applicable water quality standards. The problem is that for fresh waters impacted by cyanoHABs, no scientifically-based framework exists for making this designation. This study describes the development of a data-based framework using the Ohio waters of western Lake Erie as an exemplar for large lakes impacted by cyanoHABs. To address this designation for Ohio's open waters, the Ohio Environmental Protection Agency (EPA) assembled a group of academic, state and federal scientists to develop a framework that would determine the criteria for Ohio EPA to consider in deciding on a recreation use impairment designation due to cyanoHAB presence. Typically, the metrics are derived from on-lake monitoring programs, but for large, dynamic lakes such as Lake Erie, using criteria based on discrete samples is problematic. However, significant advances in remote sensing allows for the estimation of cyanoHAB biomass of an entire lake. Through multiple years of validation, we developed a framework to determine lake-specific criteria for designating a waterbody as impaired by cyanoHABs on an annual basis. While the criteria reported in this manuscript are specific to Ohio's open waters, the framework used to determine them can be applied to any large lake where long-term monitoring data and satellite imagery are available.

A space-time geostatistical model for probabilistic estimation of harmful algal bloom biomass and areal extent.

Harmful algal blooms (HABs) have been increasing in intensity worldwide, including the western basin of Lake Erie. Substantial efforts have been made to track these blooms using in situ sampling and remote sensing. However, such measurements do not fully capture HAB spatial and temporal dynamics due to the limitations of discrete shipboard sampling over large areas and the effects of clouds and winds on remote sensing estimates. To address these limitations, we develop a space-time geostatistical modeling framework for estimating HAB intensity and extent using chlorophyll a data sampled during the HAB season (June-October) from 2008 to 2017 by five independent monitoring programs. Based on the Bayesian information criterion for model selection, trend variables explain bloom northerly and easterly expansion from Maumee Bay, wind effects over depth, and variability among sampling methods. Cross validation results demonstrate that space-time kriging explains over half of the variability in daily, location-specific chlorophyll observations, on average. Conditional simulations provide, for the first time, comprehensive estimates of overall bloom biomass (based on depth-integrated concentrations) and surface areal extent with quantified uncertainties. These new estimates are contrasted with previous Lake Erie HAB monitoring studies, and deviations among estimates are explored and discussed. Overall, results highlight the importance of maintaining sufficient monitoring coverage to capture bloom dynamics, as well as the benefits of the proposed approach for synthesizing data from multiple monitoring programs to improve estimation accuracy while reducing uncertainty.

Accuracy of data buoys for measurement of cyanobacteria, chlorophyll, and turbidity in a large lake (Lake Erie, North America): implications for estimation of cyanobacterial bloom parameters from water quality sonde measurements.

Microcystin (MCY)-producing harmful cyanobacterial blooms (cHABs) are an annual occurrence in Lake Erie, and buoys equipped with water quality sondes have been deployed to help researchers and resource managers track cHABs. The objective of this study was to determine how well water quality sondes attached to buoys measure total algae and cyanobacterial biomass and water turbidity. Water samples were collected next to two data buoys in western Lake Erie (near Gibraltar Island and in the Sandusky subbasin) throughout summers 2015, 2016, and 2017 to determine correlations between buoy sonde data and water sample data. MCY and nutrient concentrations were also measured. Significant (P < 0.001) linear relationships (R2 > 0.75) occurred between cyanobacteria buoy and water sample data at the Gibraltar buoy, but not at the Sandusky buoy; however, the coefficients at the Gibraltar buoy differed significantly across years. There was a significant correlation between buoy and water sample total chlorophyll data at both buoys, but the coefficient varied considerably between buoys and among years. Total MCY concentrations at the Gibraltar buoy followed similar temporal patterns as buoy and water sample cyanobacterial biomass data, and the ratio of MCY to cyanobacteria-chlorophyll decreased with decreased ambient nitrate concentrations. These results suggest that buoy data are difficult to compare across time and space. Additionally, the inclusion of nitrate concentration data can lead to more robust predictions on the relative toxicity of blooms. Overall, deployed buoys with sondes that are routinely cleaned and calibrated can track relative cyanobacteria abundance and be used as an early warning system for potentially toxic blooms.

Interactions between nitrogen form, loading rate, and light intensity on Microcystis and Planktothrix growth and microcystin production.

The toxin-producing, bloom-forming cyanobacterial genera Microcystis and Planktothrix require fixed nitrogen (N), such as nitrate, ammonium, or organic N (e.g., urea) for growth and production of microcystins (MC). Bioavailable N can enter lakes in pulses via tributary discharge and through in-lake recycling, which can maintain low N concentrations. Additionally, light intensity has been suggested to play a role in MC production. This study examined how three forms of N (nitrate, ammonium, and urea) interacted with N loading rate (one large pulse vs. many small pulses) and light intensity to stimulate Microcystis and Planktothrix growth and MC production using nutrient enrichment experiments. Enrichments of nitrate, ammonium, and urea resulted in greater cyanobacterial biovolumes and MC concentrations than phosphorus-only enrichments, and there was no difference between pulse (100 µmol/L) and press treatments (8.3 µmol/L every 4 h). Analysis of mcyD transcripts showed significant up-regulation within 4 h of ammonium and urea enrichment. High light intensities (300 µmol photons/m2/s) with N enrichment resulted in greater cyanobacterial biovolumes and MC concentrations than lower light intensities (30 and 3 µmol photons/m2/s). Overall, the results suggest Microcystis and Planktothrix can use many forms of N and that high light intensities enhance MC production during elevated N concentrations. Moreover, the results here further demonstrate the importance of considering N, as well as P, in management strategies aimed at mitigating cyanobacterial blooms.