There have been sporadic reports of aquatic, benthic Microcoleus proliferations in freshwater rivers, lakes, and reservoirs for four decades, with reports increasing in frequency over the last twenty years, suggesting a possible rise in their global distribution, frequency, and intensity. Microcoleus can produce anatoxins which are neurotoxic, and ingestion of toxic mats has caused hundreds of dog fatalities and raised serious human and ecological health concerns. This review synthesizes and evaluates current knowledge on Microcoleus distribution, taxonomy, toxin production, toxicity, ecology, environmental drivers, and biotic interactions. Toxin-producing Microcoleus have been reported in at least 18 countries, though many regions have not conducted toxin testing, suggesting a broader but under-reported distribution. Proliferations occur across diverse habitats, including cobble-bedded streams, large sandy rivers, reservoirs, and lakes. Microcoleus proliferations also occur on macrophytes, both in lakes and rivers. Genomic analyses currently classify anatoxin-producing Microcoleus into distinct species, with all known anatoxin-producers isolated from freshwater ecosystems. Anatoxin concentrations vary widely over space and time, within and among waterbodies. While studies on environmental drivers remain limited, research in cobble-bedded rivers suggests that moderate enrichment of dissolved inorganic nitrogen and low dissolved reactive phosphorus concentrations in the water column promote proliferation. Metagenomic approaches have revealed unique nutrient acquisition and storage strategies used by Microcoleus. Key knowledge gaps remain around the environmental and ecological triggers of proliferation, toxin production, genomic diversity and microbial interactions. Addressing these gaps through coordinated, global studies using robust datasets and consistent methods is critical to improve prediction, monitoring, and mitigation of this increasingly widespread public and ecological health threat.

World Health Organization (WHO) guidance on the management of cyanobacteria in recreational waters provides a comprehensive framework for reducing human health risks. To provide a global overview of how cyanobacterial risks in recreational waters are managed and to better understand capacity gaps and best practice, we surveyed expert opinion across the world. A total of 161 survey responses were collected from 49 countries. Results indicate widespread variation in the extent of recreational water monitoring. Routine monitoring often varies by region in larger, more populous countries, while seasonal monitoring, typically conducted at variable intervals, was the most used approach. Approximately half of the countries reported either a lack of routine monitoring or a combination of different monitoring patterns. Notably, whereas nearly all countries reported methods for planktonic sampling, only one-third of countries reported methods for benthic sampling. Globally, we conclude that the assessment and management of benthic cyanobacteria risks lag behind that of planktonic cyanobacteria risks. Cell counts and pigments were the most common assessment techniques for planktonic cyanobacteria. Microcystis and Oscillatoria were the most frequently reported problematic genus of planktonic and benthic cyanobacteria, respectively, and microcystins were the most frequently reported cyanotoxins of concern. A wide variety of management responses and risk communication methods were reported with the primary action trigger being the presence of cyanobacterial scums. Experts across the world were mostly unsatisfied with recreational water risk management regimes, with over 80

β-N-Methylamino-l-alanine (BMAA), a neurotoxic nonproteinogenic amino acid implicated in the development of neurodegenerative diseases, has been broadly associated with cyanobacteria and bioaccumulation through the food web. Early high-level detections, from poorly selective high-performance liquid chromatography-fluorescence detection (HPLC-FLD) methods, have never been repeated using modern, selective tandem mass spectrometry methods. We developed a hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method for total BMAA analysis, using an improved solid-phase extraction sample cleanup to reduce suppression. We identified oxidative degradation products of BMAA, formed during strong acid hydrolysis, due to residual nitrate in culture media and implemented a dewatering step to resolve this problem. The method was validated using Spirulina powder, environmental, and two cultured cyanobacterial samples, achieving a reporting limit of 0.3 mg/kg in dry cyanobacterial samples. The validated method was then applied to an additional 30 cyanobacteria cultures from Aotearoa New Zealand. BMAA was only detected in one strain, Planktothrix sp. CAWBG-35, at 2.3 mg/kg. These results indicate that BMAA is not produced at the high levels originally reported (e.g., g/kg) in freshwater cyanobacteria and suggest that BMAA production is not widespread among freshwater cyanobacteria from Aotearoa New Zealand. These results have broad implications for future risk-based environmental and public health monitoring.

Nodularin is a cyanobacterial toxin predominantly produced by Nodularia spumigena Mertens. This cyanobacterium has been associated with fish, bird, and animal deaths, and the toxin can contaminate fish and shellfish intended for human consumption. With climate change, blooms of Nodularia spumigena are becoming more frequent and more widespread, meaning that the risk this cyanobacterium poses to human health must be evaluated. However, toxicity information is scarce, meaning that data from the closely related toxin class, microcystins, are currently used instead. In the current study, the LD50 of nodularin-R by intraperitoneal injection was found to be 50 µg/kg, consistent with previous reports. For the first time, the LD50 of nodularin-R by oral administration was determined using the voluntary consumption of laced cream cheese (7.5 mg/kg), which is similar to that reported for microcystin-LR (10.9 mg/kg). The use of microcystin toxicology data in food safety assessments for nodularin, therefore, seems reasonable—but it is critical that an oral repeated-dose study is performed on nodularin to better understand the risk posed by this toxin through chronic exposure.

This study reports the first documented accumulation of lyngbyatoxin-a (LTA), a cyanotoxin produced by marine benthic cyanobacteria, in edible shellfish in Aotearoa New Zealand. The study investigates two bloom events in 2022 and 2023 on Waiheke Island, where hundreds of tonnes of marine benthic cyanobacterial mats (mBCMs) washed ashore each summer. Genetic analysis identified the cyanobacterium responsible for the blooms as Okeania sp., a genus typically found in tropical marine ecosystems. Analysis by liquid chromatography–tandem mass spectrometry indicated that the cyanobacteria produced a potent dermatoxin, lyngbyatoxin-a (LTA), and that LTA had accumulated in marine snails, rock oysters and cockles collected near the mats. Snails contained the highest levels of LTA (up to 10,500 µg kg−1). The study also demonstrated that the LTA concentration was stable in composted mats for several months. The presence of LTA in edible species and its stability over time raise concerns about the potential health risks to humans consuming LTA-contaminated seafood. This underlines the need for further studies assessing the risks of human exposure to LTA through seafood consumption, particularly as climate change and eutrophication are expected to increase the frequency of mBCM blooms. The study highlights the need to develop public health risk management strategies for mBCMs.

The genus Gambierdiscus produces an array of bioactive hydrophilic and lipophilic secondary metabolites that range in mode of action and toxicity. In this study, the metabolite fingerprint was mapped for thirteen Gambierdiscus, five Coolia and two Fukuyoa species (34 isolates) by assessing the production of 56 characterised secondary metabolites. Gambierdiscus polynesiensis was the only species to produce Pacific-ciguatoxin-3B (P-CTX3B), P-CTX3C, iso-P-CTX3B/C, P-CTX4A, P-CTX4B and iso-P-CTX4A/B. G. australes produced maitotoxin-1 (MTX-1) and MTX-5, G. cheloniae produced MTX-6 and G. honu produced MTX-7. Ubiquitous production of 44-methylgambierone was observed amongst all the Gambierdiscus isolates, with nine species also producing gambierone. Additional gambierone analogues, including anhydrogambierone (tentatively described herein), were also detected in all Gambierdiscus species, two Coolia and two Fukuyoa species. Gambieroxide was detected in G. lewisii and G. pacificus and gambieric acid A was detected in ten Gambierdiscus species, with G. australes (CAWD381) being the only isolate to produce gambieric acids A–D. This study has demonstrated that the isolates tested to date produce the known CTXs or MTXs, but not both, and highlighted several species that produced ‘unknown’ compounds displaying characteristics of cyclic polyethers, which will be the focus of future compound discovery efforts.

Cyanobacterial blooms are increasing in frequency and intensity globally, impacting lake ecosystem health and posing a risk to human and animal health due to the toxins they can produce. Cyanobacterial pigments preserved in lake sediments provide a useful means of understanding the changes that have led to cyanobacterial blooms in lakes. However, there is some uncertainty as to whether specific carotenoids are unique to certain genera or types of cyanobacteria. To fill this knowledge gap, we analyzed pigments in 34 cyanobacteria cultures and applied the findings to sediments from three New Zealand lakes. The cyanobacterial carotenoids canthaxanthin, echinenone and zeaxanthin were detected in all cultures, whereas myxoxanthophyll was only detected in ten cultures (Microcoleus, Planktothrix and the picocyanobacteria cultures; Synechococcaceae). The sum of the individual carotenoid concentrations provided the strongest relationship with cyanobacterial biomass (R2 = 0.58) and could be used in paleolimnology studies to evaluate general cyanobacterial abundance. Ratios of canthaxanthin, zeaxanthin and myxoxanthophyll relative to echinenone indicated that carotenoid ratios could be used to differentiate picocyanobacteria and bloom-forming cyanobacteria, to some degree. High zeaxanthin/echinenone ratios were measured in picocyanobacteria and low zeaxanthin/echinenone ratios were measured in bloom-forming cyanobacteria. The zeaxanthin/echinenone ratio was applied to sediment core samples where the cyanobacterial community was also evaluated by 16S rRNA gene metabarcoding, with the zeaxanthin/echine-none ratios showing similar patterns to those observed in the cultures. The preliminary assessment described here suggests that zeaxanthin/echinenone ratios could provide a valuable paleoecological proxy for evaluating historical shifts in cyanobacterial communities and warrants further exploration.

Lake ecosystems around the world are subject to multiple anthropogenic pressures leading to increased eutrophication and degraded ecosystems. The understanding of lake health at national scales is constrained by limited data, however, the increased long-term monitoring required to address such data deficiencies would be impractical, expensive and subject to significant time lags. More efficient methods of assessing contemporary water quality are needed. Lake sediments and the flux of constituents across the sediment-water interface in response to various biogeochemical cycles can result in the surficial sediments providing a time-integrated record of environmental conditions in the lake. As such, sediment geochemistry may offer a valuable and efficient indicator of contemporary lake water quality. To assess the potential of sediment indicators of water quality, geochemistry was analysed in surficial sediment samples (0-2 cm) collected from 101 lakes across New Zealand for which long-term water quality monitoring data was available. The selected lakes spanned various gradients, including lake type, trophic state, depth, latitude, altitude, catchment land use and sediment geochemistry. Linear modelling was undertaken to predict trophic state from sediment geochemistry and lake physiographic data separately and then from the two datasets combined. The combined model proved to be the strongest predictor of contemporary water quality (R-2 = 0.80) and is referred to here as the Sediment Geochemistry Trophic Model (SGTM). This model was then used to predict the trophic level in 76 unmonitored lakes. The sediment geochemistry analyses conducted for the development of the SGTM are routinely conducted for lake management projects and, aside from their predictive power for trophic levels, provide useful insights into nutrient cycling and lake restoration planning. The relationship between lake water quality and sediment geochemistry is complex, however, the SGTM offers a highly efficient method for assessing the state and drivers of contemporary water quality in unmonitored lakes.

Remote sensing using satellite imagery has been promoted as a method to broaden the scale and frequency of cyanobacterial monitoring. This relies on the ability to establish relationships between the reflectance spectra of water bodies and the abundance of cyanobacteria. A challenge to achieving this comes from a limited understanding of the extent to which the optical properties of cyanobacteria vary according to their physiological state and growth environment. The aim of the present study was to determine how growth stage, nutrient status and irradiance affect pigment concentrations and absorption spectra in two common bloom forming cyanobacterial taxa: Dolichospermum lemmermannii and Microcystis aeruginosa. Each species was grown in laboratory batch culture under a full factorial design of low or high light intensity and low, medium, or high nitrate concentrations. Absorption spectra, pigment concentrations and cell density were measured throughout the growth phases. The absorption spectra were all highly distinguishable from each other, with greater interspecific than intraspecific differences, indicating that both D. lemmermannii and M. aeruginosa can be readily differentiated using hyperspectral absorption spectra. Despite this, each species exhibited different responses in the per-cell pigment concentrations with varying light intensity and nitrate exposure. Variability among treatments was considerably higher in D. lemmermannii than in M. aeruginosa, which exhibited smaller changes in pigment concentrations among the treatments. These results highlight the need to understand the physiology of the cyanobacteria and to take caution when estimating biovolumes from reflectance spectra when species composition and growth stage are unknown.

Introduced trout can induce trophic cascades, however, a lack of pre-introduction data limits knowledge on their impact in many lakes. Traditional paleolimnological approaches have been used to study historic species changes, but until recently these have been restricted to taxa with preservable body-parts. To explore the ecosystem effects of Salmo trutta (brown trout) introduction on an oligotrophic lake in Aotearoa-New Zealand, we used a multi-marker sedimentary environmental DNA (sedDNA) approach coupled with pigments to detect changes across multiple trophic levels. DNA was extracted from core depths capturing approximately 100 years before and after the expected arrival of S. trutta, and metabarcoding was undertaken with four primer sets targeting the 12S rRNA (fish), 18S rRNA (eukaryotes) and cytochrome c oxidase (COI; eukaryotes) genes. The earliest detection of S. trutta eDNA was 1906 (1892–1919 CE with 95% high probability density function) suggesting their introduction was shortly before this. Native fish diversity (12S and 18S rRNA) decreased after the detection of S. trutta, albeit the data was patchy. A shift in overall eukaryotic and algal communities (18S rRNA and COI) was observed around 1856 (1841–1871 CE) to 1891 (1877–1904 CE), which aligns with the expected S. trutta introduction. However, taxonomy could not be assigned to many of the 18S rRNA and COI sequences. Pigment concentrations did not change markedly after S. trutta introduction. SedDNA provides a new tool for understanding the impact of disturbances such as the introduction of non-native species; however, there are still several methodological challenges to overcome.

Identifying compounds responsible for the observed toxicity of the Gambierdiscus species is a critical step to ascertaining whether they contribute to ciguatera poisoning. Macroalgae samples were collected during research expeditions to Rarotonga (Cook Islands) and North Meyer Island (Kermadec Islands), from which two new Gambierdiscus species were characterized, G. cheloniae CAWD232 and G. honu CAWD242. Previous chemical and toxicological investigations of these species demonstrated that they did not produce the routinely monitored Pacific ciguatoxins nor maitotoxin-1 (MTX-1), yet were highly toxic to mice via intraperitoneal (i.p.) injection. Bioassay-guided fractionation of methanolic extracts, incorporating wet chemistry and chromatographic techniques, was used to isolate two new MTX analogs; MTX-6 from G. cheloniae CAWD232 and MTX-7 from G. honu CAWD242. Structural characterization of the new MTX analogs used a combination of analytical chemistry techniques, including LC–MS, LC–MS/MS, HR–MS, oxidative cleavage and reduction, and NMR spectroscopy. A substantial portion of the MTX-7 structure was elucidated, and (to a lesser extent) that of MTX-6. Key differences from MTX-1 included monosulfation, additional hydroxyl groups, an extra double bond, and in the case of MTX-7, an additional methyl group. To date, this is the most extensive structural characterization performed on an MTX analog since the complete structure of MTX-1 was published in 1993. MTX-7 was extremely toxic to mice via i.p. injection (LD50 of 0.235 µg/kg), although no toxicity was observed at the highest dose rate via oral administration (155.8 µg/kg). Future research is required to investigate the bioaccumulation and likely biotransformation of the MTX analogs in the marine food web.

Lakes are becoming degraded at an accelerating rate owing to human activity, and understanding their past ecology is necessary for lake management and rehabilitation. Palaeolimnology provides numerous methods that enable the historical state of lakes to be determined. New Zealand provides an ideal setting in which to do this as human modification of the landscape occurred later here than in most regions of the world (approx. 1300 CE). Lake Oporoa is a shallow lake that is highly significant to the local indigenous Maori community. This study used multiple proxy palaeolimnology to explore how lake ecology shifted following Maori and European settlement in the catchment, and how palaeolimnological data can be used to inform lake rehabilitation and conservation measures, alongside the desires of the indigenous community. Sedimentary pollen, diatoms, bacterial communities, and elemental and hyperspectral imaging scanning were used to infer ecological changes in the lake and catchment from pre-human times to present. Following Maori settlement (approx. 1620 CE) there was gradual vegetation change and a rapid shift in diatom and bacterial assemblages, but not in phytoplankton pigments or sediment geochemistry. An increasing abundance of diatom taxa Discostella stelligera and Staurosirella cf. ovata indicates early nutrient enrichment. European pastoralism from approximately 1840 CE resulted in further deforestation, and all proxies show evidence for enhanced primary productivity driven by a combination of nutrient enrichment and changing lake levels, particularly since the 1960s. This has caused degradation in water quality and is likely to have contributed to the decline in populations of tuna (eel, Anguilla spp.). Conversations with local Maori, together with the palaeolimnological results, indicate that a culturally acceptable and realistic rehabilitation target for Lake Oporoa aligns with ecological conditions in the 1950s. The palaeoecological data provide information to guide catchment and lake revegetation and other methods of nutrient abatement, with the eventual aim of restoring culturally important tuna and native fish populations.

Freshwater picocyanobacteria form the base of microbial food webs in many lakes worldwide but have received less attention than other phytoplankton. Little is known about their potential response to environmental changes such as increased nutrient loading and climate change, due partly to the lack of available cultured and sequenced strains. Here, we isolated 25 monoclonal picocyanobacterial strains from 6 New Zealand lakes with contrasting trophic states. The use of MLA medium instead of BG11 proved highly successful for the rapid isolation of picocyanobacteria. Strains were characterised by sequencing of the 16S ribosomal RNA gene, spectrophotometry, and high-performance liquid chromatography. 16S rRNA gene analysis placed most strains within the cluster 5 picocyanobacterial lineage (sub-cluster 5.2, family: Synechococcaceae). Phylogenetic analysis showed that 12 isolates from Lakes Wakatipu, Hayes, Johnson, and Ellesmere/Te Waihora clustered with strains from a range of Northern Hemisphere locations, suggesting global dispersal of these strains. Pigment characterisation revealed that pink and brown cultures from oligotrophic and some eutrophic lakes were rich in phycoerythrin, while green cultures from eutrophic and hypertrophic lakes were rich in phycocyanin. This diverse group of freshwater cluster 5 picocyanobacterial cultures will provide a new resource to study how these critically important microbes function and respond to changing environmental stressors.

Interactions among multiple stressors, legacies of past perturbations, and the lack of historical information make it difficult to determine the influence of individual anthropogenic impacts on lakes and separate them from natural ecosystem variability. In the present study, we coupled paleolimnological approaches, historical data, and ecological experiments to disentangle the impacts of multiple long-term stressors on lake ecosystem structure and function. We found that the lake structure and function remained resistant to the impacts of catchment deforestation and erosion, and the introduction of several exotic fish species. Changes in ecosystem structure and function were consistent, with nutrient enrichment being the primary driver of change. Significant and sustained changes in the lake diatom community structure (and their nutrient requirements), bacterial community function, and paleolimnological proxies of ecosystem function coincided with nitrogen and phosphorus fertilizers in the catchment. The results highlight that the effects of increased nutrient inputs are much stronger than the influence of other, potentially significant, drivers of ecosystem change, and that the degree of nutrient impact can be underestimated by environmental monitoring due to its diffuse and accumulative nature. Delineating the effects of multiple anthropogenic drivers requires long-term records of both impacts and lake ecosystem change across multiple trophic levels.

Lakes and their catchments have been subjected to centuries to millennia of exploitation by humans. Efficient monitoring methods are required to promote proactive protection and management. Traditional monitoring is time consuming and expensive, which limits the number of lakes monitored. Lake surface sediments provide a temporally integrated representation of environmental conditions and contain high microbial biomass. Based on these attributes, we hypothesized that bacteria associated with lake trophic states could be identified and used to develop an index that would not be confounded by non-nutrient stressor gradients. Metabarcoding (16S rRNA gene) was used to assess bacterial communities present in surface sediments from 259 non-saline lakes in New Zealand encompassing a range of trophic states from alpine microtrophic lakes to lowland hypertrophic lakes. A subset of lakes (n = 96) with monitoring data was used to identify indicator amplicon sequence variants (ASVs) associated with different trophic states. A total of 10,888 indicator taxa were identified and used to develop a Sediment Bacterial Trophic Index (SBTI), which signficantly correlated (r2 = 0.842, P < 0.001) with the Trophic Lake Index. The SBTI was then derived for the remaining 163 lakes, providing new knowledge of the trophic state of these unmonitored lakes. This new, robust DNA-based tool provides a rapid and cost-effective method that will allow a greater number of lakes to be monitored and more effectively managed in New Zealand and globally. The SBTI could also be applied in a paleolimnological context to investigate changes in trophic status over centuries to millennia.

Understanding the historical onset of cyanobacterial blooms in freshwater bodies can help identify their potential drivers. Lake sediments are historical archives, containing information on what has occurred in and around lakes over time. Paleolimnology explores these records using a variety of techniques, but choosing the most appropriate method can be challenging. We compared results obtained from a droplet digital PCR assay targeting a cyanobacterial-specific region of the 16S rRNA gene in sedimentary DNA and cyanobacterial pigments (canthaxanthin, echinenone, myxoxanthophyll and zeaxanthin) analysed using high-performance liquid chromatography in four sediment cores. There were strong positive relationships between the 16S rRNA gene copy concentrations and individual pigment concentrations, but relationships differed among lakes and sediment core depths within lakes. The relationships were more consistent when all pigments were summed, which we attribute to different cyanobacteria species, in different lakes, at different times producing different suites of pigments. Each method had benefits and limitations, which should be taken into consideration during method selection and when interpreting paleolimnological data. We recommend this biphasic approach when making inferences about changes in the entire cyanobacterial community because they yielded complementary information. Our results support the view that molecular methods can yield results similar to traditional paleolimnological proxies when caveats are adequately addressed.

Phytoplankton (including cyanobacteria) are a natural component of lake ecosystems and are the base of many food webs. However, changes in the lake catchment, the lake itself and the wider environment (e.g., climate change) can alter the composition of phytoplankton communities. Of recent concern is the increase in the abundance of cyanobacteria and the formation of blooms in many of New Zealand’s low-land lakes (>30% of those that are monitored). Because regular monitoring data does not stretch back more than 10-30 years and many lakes are not regularly monitored, it is difficult to ascertain whether the intensity of present-day blooms are a new phenomenon or are part of a pre-existing cycle. The pigments produced by cyanobacteria (and other phytoplankton) are deposited in lake sediment and can be extracted from sediment cores and surface sediment samples. Analysis of these pigments by high-performance liquid chromatography has allowed us to reconstruct the historical phytoplankton community from nine New Zealand lakes and to track the emergence of cyanobacteria in impacted lakes. We will present data on the performance of different cyanobacteria pigment indicators and several case studies to demonstrate how fossilised pigment data can be used to understand shifts in lake phytoplankton communities.

Understanding the toxicity and production rates of the various secondary metabolites produced by Gambierdiscus and cohabitating benthic dinoflagellates is essential to unravelling the complexities associated with ciguatera poisoning. In the present study, a sulphated cyclic polyether, gambierone, was purified from Gambierdiscus cheloniae CAWD232 and its acute toxicity was determined using intraperitoneal injection into mice. It was shown to be of low toxicity with an LD50 of 2.4 mg/kg, 9600 times less toxic than the commonly implicated Pacific ciguatoxin-1B, indicating it is unlikely to play a role in ciguatera poisoning. In addition, the production of gambierone and 44-methylgambierone was assessed from 20 isolates of ten Gambierdiscus, two Coolia and two Fukuyoa species using quantitative liquid chromatography–tandem mass spectrometry. Gambierone was produced by seven Gambierdiscus species, ranging from 1 to 87 pg/cell, and one species from each of the genera Coolia and Fukuyoa, ranging from 2 to 17 pg/cell. The production of 44-methylgambierone ranged from 5 to 270 pg/cell and was ubiquitous to all Gambierdiscus species tested, as well as both species of Coolia and Fukuyoa. The relative production ratio of these two secondary metabolites revealed that only two species produced more gambierone, G. carpenteri CAWD237 and G. cheloniae CAWD232. This represents the first report of gambierone acute toxicity and production by these cohabitating benthic dinoflagellate species. While these results demonstrate that gambierones are unlikely to pose a risk to human health, further research is required to understand if they bioaccumulate in the marine food web.

Microcystis is a bloom-forming genus of cyanobacteria with some genotypes that produce highly toxic microcystin hepatotoxins. In waterbodies where biological and physical factors are relatively homogenous, toxin quotas (the average amount of toxin per cell), at a single point in time, are expected to be relatively constant. In this study we challenged this assumption by investigating the spatial distribution of microcystin quotas at a single point in time on two separate occasions in a lake with a major Microcystis bloom. Microcystis cell concentrations varied widely across the lake on both sampling occasions (730- and 137-fold) together with microcystin quotas (148- and 362-fold). Cell concentrations and microcystin quotas were strongly positively correlated (R2 = 0.89, P < 0.001, n = 28; R2 = 0.67, P < 0.001, n = 25). Analysis of Microcystis strains using high-throughput sequencing of the 16S-23S rRNA intergenic spacer region showed no relationship between microcystin quota and the relative abundance of specific sequences. Collectively, the results of this study indicate an association between microcystin production and cell density that magnifies the potential for bloom toxicity at elevated cell concentrations.