Cyanotoxins occur in rivers worldwide but are understudied in lotic ecosystems relative to lakes and reservoirs. We sampled 11 large river sites located throughout the United States during June–September 2017 to determine the occurrence of cyanobacteria with known cyanotoxin-producing strains, cyanotoxin synthetase genes, and cyanotoxins. Chlorophyll a concentrations ranged from oligotrophic to eutrophic (0.5–64.4 µg L⁻¹). Cyanobacteria were present in the algal communities of all rivers (82% of samples, n = 50) but rarely dominated the phytoplankton (0–52% of total abundance; mean = 8.8%). Pseudanabaena and Planktothrix occurred most often, and many (64%) of the cyanobacterial genera identified (n = 25) have known cyanotoxin-producing strains. Cyanotoxin synthetase genes occurred in all but one river. The mcyE and sxtA genes were most common, present in 73% of rivers and 44% and 40% of samples, respectively. The cyrA gene was less common (22% of samples) but occurred in 64% of rivers. The anaC gene was detected in one river (4% of samples). Anatoxin-a and microcystins were detected at low levels (0.10–0.38 µg L⁻¹) in 2 midcontinent rivers. Cylindrospermopsins and saxitoxins were not detected. Cyanobacteria, cyanotoxin synthetase genes, and cyanotoxins were present at low concentrations throughout this subset of US rivers. Eutrophic rivers located in the midcontinent region of the United States had the highest algal biomass, abundance of cyanotoxin synthetase genes, and cyanotoxin occurrence.

Cyanobacteria are the most common toxigenic algae in inland waters. Their toxins can affect the health of aquatic and terrestrial organisms, including humans. Other algal groups, such as haptophytes (e.g., Prymnesium parvum) and euglenoids (e.g., Euglena sanguinea), can also form harmful algal blooms (HABs) whose toxins cause injury to aquatic biota but currently have no known effects on human health. Prymnesium parvum, however, is responsible for some of the worst HAB-related ecological disasters recorded in inland waters. Here, we provide an overview of the primary toxigenic algae found in U.S. inland waters: cyanobacteria (planktonic forms), P. parvum, and E. sanguinea with the objective of describing their similarities and differences in the areas of HAB ecology, algal toxins, and the potential for future range expansion of HABs. A detailed account of bloom habitats and their known associations with land cover and use is provided from the perspective of water quality. This review revealed that salinity may have an influence on inland cyanobacterial blooms and cyanotoxins that had not been fully recognized previously.

Lake Okeechobee is one of the largest freshwater lakes in the United States. As a eutrophic lake, it has frequent algal blooms composed predominantly of the cyanobacterium genus Microcystis. Many of the algal blooms are associated with the resuspension of a thixotropic benthic mud containing legacy nutrients. Since Lake Okeechobee has an area of 1732 km² (40–50 km radius) and a mean depth of only 2.7 m, there is sufficient fetch and shallow water depth to allow frequent wind, wave, and current generated events, which cause sediment resuspension. Three types of mud exist in the lake including an immobile dark-colored, consolidated mud, a brownish-colored mud, which is poorly consolidated and mobile, and a dark-colored thixotropic, highly mobile mud that is a mixture of organic matter and clay-sized minerals. Altogether, these muds contain an estimated 4.6 × 10⁶ kg of total phosphorus and commensurate high amounts of labile nitrogen. The thixotropic mud covers most of the lakebed and contains the suitable nutrient ratios to trigger algal blooms. A bioassay analysis of the thixotropic mud compared to the consolidated mud showed that it produced up to 50% more nutrient mass compared to the consolidated mud. The thixotropic mud does not consolidate, thus remains mobile. The mobility is maintained by the dynamics of the algal blooms and bacterial decay of extracellular secretions (transparent exopolymer particles) that bind sediment, transfer it to the bottom, and undergo bacterial digestion causing gas emissions, thus maintaining the organic/sediment matrix in suspension. Despite major efforts to control external nutrient loading into the lake, the high frequency of algal blooms will continue until the muds bearing legacy nutrients are removed from the lake.

In an effort to simulate the survival of cyanobacteria as they are transported from Lake Okeechobee to the estuarine habitats that receive waters from the lake, a bioassay encompassing a range of salinities was performed. An overall decline in cyanobacteria health in salinity treatments greater than 18 practical salinity units (psu) was indicated by loss of cell membrane integrity based on SYTOX® Green staining, but this loss varied by the kind of cyanobacteria present. Microcystis aeruginosa was tolerant of salinities up to 18 psu; however, higher salinities caused leaking of microcystin from the cells. Dolichospermum circinale, another common bloom-former in this system, did not tolerate salinities greater than 7.5 psu. Stimulation of mucilage production was observed and is likely a mechanism used by both species to protect organism viability. At 7.5 psu, microcystin increased relative to chlorophyll-a, providing some evidence of biosynthesis when M. aeruginosa is exposed to this salinity. This study indicates that as freshwater cyanobacteria are transported to brackish and marine waters, there will be a loss of membrane integrity which will lead to the release of cellular microcystin into the surrounding waterbody. Additional research would be needed to determine the exact effect of salinity on this relationship.

The Lake Okeechobee and the Okeechobee Waterway (Lake Okeechobee, the St. Lucie Canal and River, and the Caloosahatchee River) experienced an extensive harmful algal bloom within Lake Okeechobee, the St. Lucie Canal and River and the Caloosahatchee River in 2016. In addition to the very visible bloom of the cyanobacterium Microcystis aeruginosa, several other cyanobacteria were present. These other species were less conspicuous; however, they have the potential to produce a variety of cyanotoxins, including anatoxins, cylindrospermopsins, and saxitoxins, in addition to the microcystins commonly associated with Microcystis. Some of these species were found before, during, and 2 weeks after the large Microcystis bloom and could provide a better understanding of bloom dynamics and succession. This report provides photographic documentation and taxonomic assessment of the cyanobacteria present from Lake Okeechobee and the Caloosahatchee River and St. Lucie Canal, with samples collected June 1st from the Caloosahatchee River and Lake Okeechobee and in July from the St. Lucie Canal. The majority of the images were of live organisms, allowing their natural complement of pigmentation to be captured. The report provides a digital image-based taxonomic record of the Lake Okeechobee and the Okeechobee Waterway microscopic flora. It is anticipated that these images will facilitate current and future studies on this system, such as understanding the timing of cyanobacteria blooms and their potential toxin production.

The occurrence of cyanobacterial harmful algal blooms (cyanoHABs) is related to their physical and chemical environment. However, less is known about their associated microbial interactions and processes. In this study, cyanoHABs were analyzed as a microbial ecosystem, using 1 year of 16S rRNA sequencing and 70 metagenomes collected during the bloom season from Lake Okeechobee (Florida, USA). Biogeographical patterns observed in microbial community composition and function reflected ecological zones distinct in their physical and chemical parameters that resulted in bloom “hotspots” near major lake inflows. Changes in relative abundances of taxa within multiple phyla followed increasing bloom severity. Functional pathways that correlated with increasing bloom severity encoded organic nitrogen and phosphorus utilization, storage of nutrients, exchange of genetic material, phage defense, and protection against oxidative stress, suggesting that microbial interactions may promote cyanoHAB resilience. Cyanobacterial communities were highly diverse, with picocyanobacteria ubiquitous and oftentimes most abundant, especially in the absence of blooms. The identification of novel bloom-forming cyanobacteria and genomic comparisons indicated a functionally diverse cyanobacterial community with differences in its capability to store nitrogen using cyanophycin and to defend against phage using CRISPR and restriction-modification systems. Considering blooms in the context of a microbial ecosystem and their interactions in nature, physiologies and interactions supporting the proliferation and stability of cyanoHABs are proposed, including a role for phage infection of picocyanobacteria. This study displayed the power of “-omics” to reveal important biological processes that could support the effective management and prediction of cyanoHABs. IMPORTANCE: Cyanobacterial harmful algal blooms pose a significant threat to aquatic ecosystems and human health. Although physical and chemical conditions in aquatic systems that facilitate bloom development are well studied, there are fundamental gaps in the biological understanding of the microbial ecosystem that makes a cyanobacterial bloom. High-throughput sequencing was used to determine the drivers of cyanobacteria blooms in nature. Multiple functions and interactions important to consider in cyanobacterial bloom ecology were identified. The microbial biodiversity of blooms revealed microbial functions, genomic characteristics, and interactions between cyanobacterial populations that could be involved in bloom stability and more coherently define cyanobacteria blooms. Our results highlight the importance of considering cyanobacterial blooms as a microbial ecosystem to predict, prevent, and mitigate them.