To control harmful algae blooms (HABs), methods based on natural mechanisms are now required. We investigated the effects of an algicide derived from macrophyte metabolites, namely mixtures of gallic, tetradecanoic, heptanoic, and octanoic acids (1:1:1:1 mass ratio, a total concentration of 14 mg/L), on the biomass of cyanobacteria and other plankton and the production of microcystins under experimental conditions. Two types of microcosms have been created: simple (microalgae, cyanobacteria, and zooplankton) and complex (microalgae, cyanobacteria, zooplankton, and planktivorous fish). We observed the dynamics of the phytoplankton structure, the concentrations of microcystins and chlorophyll-a, hydrochemistry, and the status of zooplankton and fish in both types of microcosms with and without algicide for one month (from 19 July to 19 August 2021). The introduction of algicide caused changes in phytoplankton structure, a drop in cyanobacterial biomass, and a decrease in the total concentration of microcystins. Surprisingly, the contributions of the most toxic microcystins (LR form) were higher in both types of microcosms exposed to algicide than in microcosms without algicide. The inhibitory effect on the cyanobacterial biomass was most significant in complex ecosystems (containing fish), while it was only observed at the end of the exposure in simple ecosystems. Not only algicide but also phytoplankton consumed by fish and zooplankton, as well as nutrient excretory activity by both consumers, seem to have impact on cyanobacterial biomass. This study found that the using chemical substances similar to macrophyte metabolites can help regulate HABs and cyanotoxins. However, the results differ depending on ecosystem type.

Lake Okeechobee is a large eutrophic, shallow, subtropical lake in south Florida, United States. Due to decades of nutrient loading and phosphorus rich sediments, the lake is eutrophic and frequently experiences cyanobacterial harmful algal blooms (cyanoHABs). In the past, surveys of the phytoplankton community structure in the lake have been conducted by morphological studies, whereas molecular based studies have been seldom employed. With increased frequency of cyanoHABs in Lake Okeechobee (e.g., 2016 and 2018 Microcystis-dominated blooms), it is imperative to determine the diversity of cyanobacterial taxa that exist within the lake and the limnological parameters that drive bloom-forming genera. A spatiotemporal study of the lake was conducted over the course of 1 year to characterize the (cyano)bacterial community structure, using 16S rRNA metabarcoding, with coincident collection of limnological parameters (e.g., nutrients, water temperature, major ions), and cyanotoxins. The objectives of this study were to elucidate spatiotemporal trends of community structure, identify drivers of community structure, and examine cyanobacteria-bacterial relationships within the lake. Results indicated that cyanobacterial communities within the lake were significantly different between the wet and dry season, but not between periods of nitrogen limitation and co-nutrient limitation. Throughout the year, the lake was primarily dominated by the picocyanobacterium Cyanobium. The bloom-forming genera Cuspidothrix, Dolichospermum, Microcystis, and Raphidiopsis were highly abundant throughout the lake and had disparate nutrient requirements and niches within the lake. Anatoxin-a, microcystins, and nodularins were detected throughout the lake across both seasons. There were no correlated (cyano)bacteria shared between the common bloom-forming cyanobacteria Dolichospermum, Microcystis, and Raphidiopsis. This study is the first of its kind to use molecular based methods to assess the cyanobacterial community structure within the lake. These data greatly improve our understanding of the cyanobacterial community structure within the lake and the physiochemical parameters which may drive the bloom-forming taxa within Lake Okeechobee.

While freshwater cyanobacteria are traditionally thought to be limited by the availability of phosphorus (P), fixed nitrogen (N) supply can promote the growth and/or toxin production of some genera. This study characterizes how growth on N2 (control), nitrate (NO3 -), ammonium (NH4 +), and urea as well as P limitation altered the growth, toxin production, N2 fixation, and gene expression of an anatoxin-a (ATX-A) - producing strain of Dolichospermum sp. 54. The transcriptomes of fixed N and P-limited cultures differed significantly from those of fixed N-deplete, P-replete (control) cultures, while the transcriptomes of P-replete cultures amended with either NH4 + or NO3 - were not significantly different relative to those of the control. Growth rates of Dolichospermum (sp. 54) were significantly higher when grown on fixed N relative to without fixed N; growth on NH4 + was also significantly greater than growth on NO3 -. NH4 + and urea significantly lowered N2 fixation and nifD gene transcript abundance relative to the control while cultures amended with NO3 - exhibited N2 fixation and nifD gene transcript abundance that was not different from the control. Cultures grown on NH4 + exhibited the lowest ATX-A content per cell and lower transcript abundance of genes associated ATX-A synthesis (ana), while the abundance of transcripts of several ana genes were highest under fixed N and P - limited conditions. The significant negative correlation between growth rate and cellular anatoxin quota as well as the significantly higher number of transcripts of ana genes in cultures deprived of fixed N and P relative to P-replete cultures amended with NH4 + suggests ATX-A was being actively synthesized under P limitation. Collectively, these findings indicate that management strategies that do not regulate fixed N loading will leave eutrophic water bodies vulnerable to more intense and toxic (due to increased biomass) blooms of Dolichospermum.

Anthropogenic activities release large amounts of nitrogen (N) and phosphorus (P) nutrients into the environment. Sources of nutrients include surface and sub-surface runoffs from agricultural practices with the application of chemical fertilizers and manure as well as combined sewer overflows (CSOs). Nutrient runoffs contribute to the eutrophication of aquatic ecosystems and enhance the growth of cyanobacteria. Precipitation is an important driving force behind the runoff of nutrients from agricultural fields into surrounding water bodies. To understand the dynamics between nutrient input, precipitation and cyanobacterial growth in Missisquoi Bay, Lake Champlain (Quebec), one location in Pike River (a major tributary into the bay) and four locations in Missisquoi Bay were monitored from April to November in 2017 and 2018. Biweekly water samples were analyzed using chemical methods and high-throughput sequencing of 16S rRNA gene amplicons. High concentrations of N and P were typically measured in April and May. Three major spikes in nutrient concentrations were observed in early and mid-summer as well as early fall, all of which were associated with intense cumulative precipitation events of 40 to 100 mm within 7 days prior to sampling. Despite the high concentrations of nutrients in the spring and early summer, the cyanobacterial blooms appeared in mid to late summer as the water temperature increased. Dolichospermum sp. was the major bloom-forming cyanobacterium during both summers. A second intense bloom event of Microcystis was also observed in the fall (October and November) for both years. Variation in the cyanobacteria population was strongly associated with inorganic and readily available fractions of N and P such as nitrites and nitrates (NOx), ammonia (NH3) and dissolved organic phosphorus (DOP). During blooms, total Kjeldahl nitrogen (TKN) and total particulate phosphorus (TPP) fractions had a substantial influence on total nitrogen (TN) and total phosphorus (TP) concentrations, respectively. The abundance of bacteria involved in the metabolism of nitrogen compared to that of phosphorus revealed the importance of nitrogen on overall microbial dynamics as well as CB formation in the bay. Our findings emphasize the combined influence of precipitation events, temperature and several bioavailable fractions of nitrogen and phosphorus on cyanobacterial bloom episodes.

Cyanobacterial harmful algal blooms (HABs) are increasing in frequency and magnitude worldwide. A number of parameters are thought to underlie HABs, including the ratio at which two key elements, nitrogen (N) and phosphorus (P) are supplied, although a predictive understanding eludes us. While the physiological importance of iron (Fe) in electron transport and N-fixation is well known, relatively little is known about its impacts on the growth of freshwater cyanobacteria. Moreover, there is growing appreciation for correlated changes in the quotas of multiple elements encompassing an organism (i.e. the ionome) when the supply of one element changes, indicating that growth differences arise from complex biochemical adjustments rather than limitation of a key anabolic process by a single element. In this study, the effects of supply N:P and Fe on the growth and ionome of Dolichospermum, a nitrogen-fixing cyanobacterium found in freshwater ecosystems, were examined. Changes in both supply N:P and Fe had significant effects on yield. Consistent with prior observations, cyanobacterial growth was higher at N:P = 20, compared to N:P = 5, and quotas of all elements decreased with growth. Yield was negatively related with the degree of imbalance between dissolved supply and intracellular concentrations of not only N and Fe, but also multiple other elements. Changes in Fe supply had a significant effect on yield in N-limited conditions (N:P = 5). Again, ionome-wide imbalances decreased yield. Together, these results indicate that attention to multiple elements encompassing the ionome of a HAB-forming taxon, and the supplies of such elements may help improve the ability to forecast blooms. Such elemental interactions may be critical as limnologists begin to appreciate the staggering variation in the supplies of such elements among lakes, and anthropogenic activities continue to alter global biogeochemical cycles.

The evaluation about the relative distribution of cyanobacterial biomass between the sediment and water column would be indispensable to understand if benthic cyanobacteria are important to cyanobacterial biomass in the water column. A separation method for the rapid quantification of benthic cyanobacteria in Lake Chaohu was developed by density-gradient centrifugation. A 24 full factorial design and response surface methodology was employed to optimize the extraction protocol. Under the optimal operating parameters including 29% Percoll solution, 30 min centrifugation time, 7200 r/min centrifugation speed, and a 1:10 ratio between the volume of sediment and Percoll solution, the recovery rate of cyanobacteria in sediment was 96.73%. Temporal and spatial variations in cyanobacterial biomass in water and sediment were investigated monthly throughout a whole year. In general, cyanobacterial biomass per square centimeter in the water column showed high spatial-temporal changes, tending to increase in February and reaching a peak in April at some sites due to the growth of Dolichospermum. The second peak arrived in July and September and was caused by the rapid growth of Microcystis. Concurrently, cyanobacteria biomass per unit area in sediment showed a clear temporal change pattern, increasing from October and reaching a peak level in February at all the sampling sites. The average ratio of cyanobacterial biomass in water to that in sediment was lowest in January at 1.48 and increased to the highest level in July at 318.61. Although Microcystis and Dolichospermum were dominant species in the water column, only Microcystis was observed in the Percoll solution extraction from sediment. Microscopic observation revealed that a very small fraction of Microcystis cells could survive in sediment, and most of the cells decomposed when the water temperature increased after June. Therefore, the contribution of the recruitment of cyanobacteria could be negligible in Lake Chaohu.

Combining coagulants with ballast (natural soil or modified clay) to remove cyanobacteria from the water column is a promising tool to mitigate nuisance blooms. Nevertheless, the possible effects of this technique on different toxin-producing cyanobacteria species have not been thoroughly investigated. This laboratory study evaluated the potential effects of the \"Floc and Sink\" technique on releasing microcystins (MC) from the precipitated biomass. A combined treatment of polyaluminium chloride (PAC) with lanthanum modified bentonite (LMB) and/or local red soil (LRS) was applied to the bloom material (mainly Dolichospermum circinalis and Microcystis aeruginosa) of a tropical reservoir. Intra and extracellular MC and biomass removal were evaluated. PAC alone was not efficient to remove the biomass, while PAC + LMB + LRS was the most efficient and removed 4.3-7.5 times more biomass than other treatments. Intracellular MC concentrations ranged between 12 and 2.180 µg L-1 independent from the biomass. PAC treatment increased extracellular MC concentrations from 3.5 to 6 times. However, when combined with ballast, extracellular MC was up to 4.2 times lower in the top of the test tubes. Nevertheless, PAC + LRS and PAC + LMB + LRS treatments showed extracellular MC concentration eight times higher than controls in the bottom. Our results showed that Floc and Sink appears to be more promising in removing cyanobacteria and extracellular MC from the water column than a sole coagulant (PAC).

The ADA clade of Nostocales cyanobacteria, a group that is prominent in current harmful algal bloom events, now includes over 40 genome sequences with the recent addition of sixteen novel sequenced genomes (Dreher et al., Harmful Algae, 2021). Fourteen genomes are complete (closed), enabling highly detailed assessments of gene content and genome architecture. ADA genomes contain 5 rRNA operons, genes expected to support a photoautotrophic and diazotrophic lifestyle, and a varied array of genes for the synthesis of bioactive secondary metabolites. Genes for the production of the taste-and-odor compound geosmin and the four major classes of cyanotoxins - anatoxin-a, cylindrospermopsin, microcystin and saxitoxin - are represented in members of the ADA clade. Notably, the gene array for the synthesis of cylindrospermopsin by Dolichospermum sp. DET69 was located on a plasmid, raising the possibility of facile horizontal transmission. However, genes supporting independent conjugative transfer of this plasmid are lacking. Further, analysis of genomic loci containing this and other cyanotoxin gene arrays shows evidence that these arrays have long-term stability and do not appear to be genomic islands easily capable of horizontal transmission to other cells. There is considerable diversity in the gene complements of individual ADA genomes, including the variable presence of physiologically important genes: genomes in three species-level subclades lack the gas vesicle genes that facilitate a planktonic lifestyle, and, surprisingly, the genome of Cuspidothrix issatschenkoi CHARLIE-1, a reported diazotroph, lacks the genes for nitrogen fixation. Notably, phylogenetically related genomes possess limited synteny, indicating a prominent role for chromosome rearrangements during ADA strain evolution. The genomes contain abundant insertion sequences and repetitive transposase genes, which could be the main drivers of genome rearrangement through active transposition and homologous recombination. No prophages were found, and no evidence of viral infection was observed in the bloom population samples from which the genomes discussed here were derived. Phages thus seem to have a limited influence on ADA evolution.

The genome sequences of 16 Nostocales cyanobacteria have been determined. Most of them are complete or near-complete genome sequences derived by long-read metagenome sequencing of recent harmful algal blooms (HABs) in freshwater lakes without the potential bias of culture isolation. The genomes are all members of the recently recognized ADA clade (Driscoll et al., Harmful Algae, 77:93, 2018), which we argue represents a genus. We identify 10 putative species-level branches within the clade, on the basis of 91-gene phylogenomic and average nucleotide identity analyses. The assembled genomes each correspond to a single morphotype in the original sample, but distinct genomes from different HABs in some cases correspond to similar morphotypes. We present data indicating that the ADA clade is a highly significant component of current cyanobacterial HABs, including members assigned to the prevalent Dolichospermum and Aphanizomenon genera, as well as Cuspidothrix and Anabaena. In general, currently used genus and species names within the ADA clade are not monophyletic. We infer that the morphological characters routinely used in taxonomic assignments are not reliable for discriminating species within the ADA clade. Taxonomic revisions will be needed to create a genus with a single name (we recommend Anabaena) and to adopt species names that do not depend on morphological traits that lack sufficient discrimination and specificity, while recognizing the utility of some easily observable and distinct morphologies.

Harmful algal blooms caused by cyanobacteria have been increasing in frequency worldwide. However, the main environmental drivers of this change are often difficult to identify because of the effects of the interaction between eutrophication and climate change. Recently, filamentous N2-fixing cyanobacteria and non-diazotrophic Microcystis have been observed to be co-existing and undergoing succession in some eutrophic lakes. However, the succession patterns of dominant cyanobacteria and the factors driving this in mesotrophic lakes are not well understood. We hypothesized that the changes in cyanobacterial assemblages in mesotrophic lakes could result in a relatively high risks of toxic blooms, and that these changes are associated with the global climatic changes. We tested these hypotheses using data from the subtropical mesotrophic Lake Erhai. We found that the high spatiotemporal variability in the cyanobacterial community, and the increase in biomass were driven primarily by the growth of bloom-forming cyanobacterial taxa. Species-specific biomasses were related to a different environmental stressor; increases in dissolved organic carbon (DOC) concentrations were statistically associated with an increase of Microcystis biomass, whereas increases in surface water temperature favored higher biomass of Pseudanabaena at low transparency and high concentration of phosphorus. In addition, low nitrogen- to- phosphorus ratios were identified as potential determinants of the abundance of N2-fixing Dolichospermum. Furthermore, changes in the concentration of DOC, total nitrogen, pH and water transparency levels were found to affect the composition of Microcystis morphotypes and genotypes mostly. This study highlights that the toxic to non-toxic Microcystis ratio might increase with the water darkening and browning (which occurs in many subtropical plateau lakes). Lake management strategies, therefore, need to consider the toxicity of cyanobacterial assemblages in mesotrophic lakes over the intensity of the cyanobacterial blooms.