Bloom-forming Microcystis Research Articles

Decoding Microcystis aeruginosa quorum sensing through AHL-mediated transcriptomic molecular regulation mechanisms

Acyl-homoserine lactone (AHL) serves as a key signaling molecule for quorum sensing (QS) in bacteria. QS-related genes and physiological processes in Microcystis aeruginosa remain elusive. In this study, we elucidated the regulatory role of AHL-mediated QS in M. aeruginosa. Using AHL activity extract and transcriptomic analysis, we revealed significant effects of the AHL on growth and photosynthesis. AHL significantly increased chlorophyll a (Chl-a) content and accelerated photosynthetic rate thereby promoting growth. Transcriptome analysis revealed that AHL stimulated the up-regulation of photosynthesis-related genes (apcABF, petE, psaBFK, psbUV, etc.) as well as nitrogen metabolism and ribosomal metabolism. In addition, AHL-regulated pathways are associated with lipopolysaccharide and phenazine synthesis. Our findings deepen the understanding of the QS system in M. aeruginosa and are important for gaining insights into the role of QS in Microcystis bloom formation. It also provides new insights into the prevalence of M. aeruginosa in water blooms.

Indole-3-acetic acid promotes growth in bloom-forming Microcystis via an antioxidant response

Interactions between bacteria and phytoplankton in the phycosphere facilitate and constrain biogeochemical cycling in aquatic ecosystems. Indole-3-acetic acid (IAA) is a bacterially produced chemical signal that promotes growth of phytoplankton and plants. Here, we explored the impact of IAA on bloom-forming cyanobacteria and their associated bacteria. Exposure to IAA and its precursor, tryptophan, resulted in a strong growth response in a bloom of the freshwater cyanobacterium, Microcystis. Metatranscriptome analysis revealed the induction of an antioxidant response in Microcystis upon exposure to IAA, potentially allowing populations to increase photosynthetic rate and overcome internally generated reactive oxygen. Our data reveal that co-occurring bacteria within the phycosphere microbiome exhibit a division of labor for supportive functions, such as nutrient mineralization and transport, vitamin synthesis, and reactive oxygen neutralization. These complex dynamics within the Microcystis phycosphere microbiome are an example of interactions within a microenvironment that can have ecosystem-scale consequences.

The extracellular polysaccharide determine the physico-chemical surface properties of Microcystis.

Microcystis possesses the capacity to form colonies and blooms in lakes and reservoirs worldwide, causing significant ecological challenges in aquatic ecosystems. However, little is known about the determining factors of physico-chemical surface properties that govern the competitive advantage of Microcystis. Here, The physico-chemical surface properties of Microcystis wesenbergii and Microcystis aeruginosa, including specific surface area (SSA), hydrophobicity, zeta potential, and functional groups were investigated. Additionally, the extracellular polysaccharide (EPS) were analyzed. Laboratory-cultured Microcystis exhibited hydrophilic, a negative zeta potential and negatively charged. Furthermore, no significant relationship was shown between these properties and the cultivation stage. Microcystis wesenbergii exhibited low free energy of cohesion, high surface free energy, high growth rate, and high EPS content during the logarithmic phase. On the other hand, M. aeruginosa displayed lower free energy of cohesion, high surface free energy, high EPS content, and high growth rate during the stationary phase. These characteristics contribute to their respective competitive advantage. Furthermore, the relationship between EPS and surface properties was investigated. The polysaccharide component of EPS primarily influenced the SSA and total surface energy of Microcystis. Likewise, the protein component of EPS influenced hydrophobicity and surface tension. The polysaccharide composition, including glucuronic acid, xylose, and fructose, mainly influenced surface properties. Additionally, hydrophilic groups such as O-H and P-O-P played a crucial role in determining hydrophobicity in Microcystis. This study elucidates that EPS influenced the SSA, hydrophobicity, and surface free energy of Microcystis cells, which in turn impact the formation of Microcystis blooms and the collection.

Isopropylthiol emission by bloom-forming Microcystis: Biochemistry, ecophysiology and semiochemistry of a volatile organosulfur compound

Microcystis species not only produce toxic cyanobacterial blooms, but can be a significant source of taste and odour. Previous studies have associated foul-smelling volatile organic sulfur compounds (VOSCs) with Microcystis blooms, but have largely attributed these compounds to bacterial bloom decomposition. However, earlier reports of the production of isopropylthio compounds by several Microcystis strains suggests that these cyanobacteria may themselves be a source of these VOSCs. Sulphur compounds have been shown to play important semiochemical roles in algal cell protection and grazer interactions in marine systems, but little is known about the production and chemical ecology of freshwater cyanobacterial VOSCs. To address this knowledge gap, we undertook the first detailed investigation of the biochemistry, ecophysiology and semiochemistry of these compounds and their production by Microcystis, and tested the hypothesis that they act as multifunctional semiochemicals in processes related to cell protection and grazer defence. Using short-term incubations and an adapted headspace-GC–MS technique, we investigated VOSC production by axenic and non-axenic strains, and verified that isopropylthio compounds are in fact produced by these cyanobacteria, identifying 5 isopropyl moiety-containing VOSCs (isopropylthiol (ISH), isopropylmethyl sulfide, isopropyl methyl disulfide, diisopropyl disulfide (ISSI) and diisopropyl trisulfide) as well as methanethiol in three strains. Further studies with the axenic strain Microcystis PCC 7806 using different light regimes, metabolic inhibitors (sodium azide, DCMU), the antioxidant enzyme catalase and stable labelled precursors (hydrogencarbonate, acetates and sulfate) demonstrated that ISH is a true exo-metabolite, synthesized via the acetate pathway. It is actively produced and continuously excreted by the cyanobacteria during growth, with minimal internal storage or post-lysis catalytic generation. The molar ratios of the redox pair ISH/ISSI are not directly involved in the photosynthetic and respiratory electron transport chains, but dependant on the redox state of the cell - likely mediated by reactive oxygen species (ROS), as shown by a marked effect of catalase. These results, along with toxicological and behavioural assays using the two aquatic invertebrates Thamnocephalus platyurus and Daphnia magna indicate that ISH plays multiple important physiological and ecological roles. It acts as an effective antioxidant against high ROS levels, as often experienced in surface blooms, it elicits avoidance-related behavioural responses in grazer communities and at high levels, it can be toxic to some invertebrates.

Dynamic response of bacterial communities to Microcystis blooms: A three-year study

Although nutrient availability is widely recognized as the driving force behind Microcystis blooms, identifying the microorganisms that play a pivotal role in their formation is a challenging task. Our understanding of the contribution of bacterial communities to the development of Microcystis blooms remains incomplete, despite the fact that the relationship between Microcystis and bacterial communities has been extensively investigated. Most studies have focused on their interaction for a single year rather than for multiple years. To determine key bacteria crucial for the formation of Microcystis blooms, we collected samples from three sites in the Daechung Reservoir (Chuso, Hoenam, and Janggye) over three years (2017, 2019, and 2020). Our results indicated that Microcystis bloom-associated bacterial communities were more conserved across stations than across years. Bacterial communities could be separated into modules corresponding to the different phases of Microcystis blooms. Dolichospermum and Aphanizomenon belonged to the same module, whereas the module of Microcystis was distinct. The microbial recurrent association network (MRAN) showed that amplicon sequence variants (ASVs) directly linked to Microcystis belonged to Pseudanabaena, Microscillaceae, Sutterellaceae, Flavobacterium, Candidatus Aquiluna, Bryobacter, and DSSD61. These ASVs were also identified as key indicators of the bloom stage, indicating that they were fundamental biological elements in the development of Microcystis blooms. Overall, our study highlights that, although bacterial communities change annually, they continue to share core ASVs that may be crucial for the formation and maintenance of Microcystis blooms.

Harmful Algal Blooms: A Prolific Issue in Urban Stormwater Ponds

Nutrient-driven cyanobacteria blooms are an increasingly common issue in freshwater environments, particularly in anthropogenically altered landscapes. As stormwater runoff is one of the largest sources of nutrients for freshwater bodies, stormwater retention ponds in urban and suburban areas are likely environments for harmful cyanobacteria blooms and were thus targeted for an in-depth investigation assessing taxonomic composition, bloom morphological composition, toxicity, and impact of nutrients and other environmental drivers. Eighty-seven algal blooms were sampled from 2019 to 2022 in the greater Wilmington, North Carolina, area. Physicochemical parameters were recorded, and blooms were classified by type (defined as surface mat, surface scum, water column distribution, or benthic mat) and dominant taxa. Blooms of potentially toxic cyanobacteria genera in the water column of stormwater retention ponds were most prevalent. Dissolved inorganic phosphorus was significantly related to chlorophyll-α, Microcystis bloom formation, and the production of microcystin. Seventeen potentially toxic cyanobacteria genera were identified in retention ponds, some of whose blooms demonstrated detectable microcystin. Monoclonal cultures isolated from some blooms were found to produce anabaenopeptin and saxitoxin. The results demonstrate a higher incidence of potentially toxic cyanobacteria over other bloom-forming taxa (chlorophytes, euglenoids, chrysophytes, dinoflagellates, and diatoms) in the 39 water bodies sampled. The frequency of blooms occurring in stormwater ponds and the diversity of potentially toxic cyanobacteria identified suggest such harmful blooms are likely widespread in similar freshwater environments across multiple urbanizing areas. The blooms sampled in this study were all within residential, commercial, or recreational areas easily accessible to people, presenting serious hazards to both environmental and public health.

The role of morphological changes in Microcystis adaptation to nutrient availability at the colonial level

Colony formation is a key trait facilitating the formation of Microcystis blooms. However, the role of morphological changes (e.g., colony size and tightness) in the adaptation to nutrient availability is not fully understood. In this study, we analyzed the morphological changes under both nutrient sufficiency and deficiency. Accordant morphological changes were found with both an isolated colonial strain and mixed field colonies. Colonies that were limited by nutrients became bloated and uncompacted structures, and this change was more pronounced under N deficiency. This looser morphology increased the availability of intra-colony light and relieved the size effect. When nutrients were sufficient, small colonies emerged, which helped to maintain rapid growth (0.32 day−1). Our study highlighted probable role of morphological variations in: (1) diminishing intra-colony self-shading when facing nutrient deficiency; and (2) enlarging the population under high trophic levels by generating daughter colonies. These roles were also verified using field data from Lake Taihu, which further indicated that the seasonal succession of morphospecies was probably the result of adaptive morphological changes. Adaptive morphological changes offer advantages against fluctuations in nutrient availability, which should be considered when attempting to restrain bloom formation.

Warming and Salt Intrusion Affect Microcystin Production in Tropical Bloom-Forming Microcystis.

The Vietnamese Mekong Delta is predicted to be one of the regions most impacted by climate change, causing increased temperature and salinity in inland waters. We hypothesized that the increase in temperature and salinity may impact the microcystin (MC) production of two Microcystis strains isolated in this region from a freshwater pond (strain MBC) and a brackish water pond (strain MTV). The Microcystis strains were grown at low (27 °C), medium (31 °C), high (35 °C) and extremely high (37 °C) temperature in flat photobioreactors (Algaemist). At each temperature, when cultures reached a stable state, sea salt was added to increase salinity to 4‰, 8‰, 12‰ and 16‰. MC concentrations and cell quota were reduced at high and extremely high temperatures. Salinity, in general, had comparable effects on MC concentrations and quota. At a salinity of 4‰ and 8‰, concentrations of MC per mL of culture and MC cell quota (based on chlorophyll, dry-weight and particle counts) were higher than at 0.5‰, while at the highest salinities (12‰ and 16‰) these were strongly reduced. Strain MBC produced five MC variants of which MC-RR and MC-LR were most abundant, followed by MC-YR and relatively low amounts of demethylated variants dmMC-RR and dmMC-LR. In strain MTV, MC-RR was most abundant, with traces of MC-YR and dmMC-RR only in cultures grown at 16‰ salinity. Overall, higher temperature led to lower MC concentrations and cell quota, low salinity seemed to promote MC production and high salinity reduced MC production. Hence, increased temperature and higher salinity could lead to less toxic Microcystis, but since these conditions might favour Microcystis over other competitors, the overall biomass gain could offset a lower toxicity.

Arsenic Accumulation and Biotransformation Affected by Nutrients (N and P) in Common Blooming-Forming Microcystis wesenbergii (Komárek) Komárek ex Komárek (Cyanobacteria)

Arsenic accumulation and biotransformation in algae was mostly carried out in a medium that contained far higher nutrient concentrations than that in natural freshwaters. The obtained results might have limited environmental validity and result in a failure to describe authentic arsenic biogeochemical cycles in natural freshwater systems. To validate the assumption, arsenic accumulation, and biotransformation in common bloom forming Microcystis wesenbergii was performed under a high nutrient concentration in BG11 medium (N = 250 mg/L, P = 7.13 mg/L), and adjusted low nutrients that mimicked values in natural freshwaters (N = 1.5 mg/L, P = 0.3 mg/L). The growth rate and maximum M. wesenbergii cell density were much lower in the high nutrient set, but more inhibition was shown with increasing ambient iAs(V) concentrations both in the high and low nutrient sets. The proportion of intracellular contents in total arsenicals decreased with increasing iAs(V) concentrations in both high and low nutrient sets but increased with incubation time. Intracellular iAs(III) was not found in the high nutrient set, while it formed high concentrations that could be comparable to that of an extracellular level in the low nutrient set. M. wesenbergii could methylate arsenic, and a higher proportion of organoarsenicals was formed in the low nutrient set. Lower intracellular MMA(V) and DMA(V) concentrations were found in the high nutrient set; contrarily, they presented a higher concentration that could be comparable to the extracellular ones in the low nutrient set. The results demonstrated that different nutrient regimes could affect arsenic accumulation and biotransformation in M. wesenbergii, and low nutrient concentrations could inhibit the excretion of iAs(III), MMA(V) and DMA(V) out of cells. Further investigations should be based on natural freshwater systems to obtain an authentic arsenic accumulation and biotransformation in cyanobacteria.

Identification and characterization of alkaline phosphatase gene phoX in Microcystis aeruginosa PCC7806.

PhoX is an extracellular alkaline phosphatase that is widely found in cyanobacteria and plays an important role in the conversion of extracellular organophosphorus into soluble inorganic phosphorus. However, the phoX gene has not yet been experimentally confirmed to exist in bloom-forming Microcystis species. In this study, we identified a putative phoX gene (GenBank accession no. ARI79942.1) in M. aeruginosa PCC7806 and overexpressed it in Escherichia coli 21 (DE3). The expressed PhoX protein displayed phosphodiesterase and phosphomonoesterase activities. In contrast to other bacterial PhoX proteins, which are activated mainly by Ca2+, Microcysits PhoX was most strongly activated by Mg2+, followed by Co2+, Ca2+, Zn2+ and Mn2+, but it was inhibited by Ni2+. Sequence analysis showed that phoX was highly conserved in the Microcystis genus (DNA similarity > 96% between species). phoX expression responded significantly to different environmental phosphorus levels. When PCC7806 cells were cultured in phosphorus-deficient medium (BG11-P), phoX expression reached its highest level at 2h and then decreased to a low level at 4h. Organophosphate induced the expression of phoX; its expression reached the highest level at 4h and was maintained at a high level at 6h. Our results confirmed a putative phoX gene and demonstrated that the phoX gene of Microcystis is conserved.

Magnetic Fe3O4 nanoparticles enhance cyanobactericidal effect of allelopathic p-hydroxybenzoic acid on Microcystis aeruginosa by enhancing hydroxyl radical production

Recently, considerable progress has been made in the environmental application of nanotechnology. However, little is known about how nanomaterials might affect the cyanobacterial suppression potential of allelochemicals. In this study, a microcosm was employed to simulate and verify the effect of magnetic Fe3O4 nanoparticles (MFN) on the inhibitory influence of allelopathic hydroxybenzoic acid (p-Ha) on bloom-forming Microcystis aeruginosa. MFN had a hormetic effect on cyanobacterial growth. At a neutral concentration of 182 mg/L, MFN enhanced the algal suppression by p-Ha and decreased the IC50 by half, which was significantly and positively associated with the amount of OH. Furthermore, adding MFN induced a stronger physiological response than treatment with only p-Ha. The cellular integrity was severely disrupted for the cyanobacterium M. aeruginosa. The total protein content decreased rapidly to inactivate the algae by limiting the amounts of extracellular microcystin and polysaccharide released. The modification of the effect of p-Ha by MFN was reflected by the intracellular NO content of M. aeruginosa. In addition, the typical radical scavengers ascorbic acid and 5,5-dimethyl-1-pyrroline N-oxide decreased OH production to weaken algal suppression under the combined treatment with p-Ha and MFN. By contrast, the addition of Fe3+ and increasing the light intensity triggered the generation of OH and strong cyanobacterial suppression. Thus, MFN could enhance the cyanobacterial control efficiency of p-Ha and decrease the input of allelochemicals in the field. These findings suggest a novel mode of allelochemical modification by nanomaterials as a promising cyanobactericide for harmful algal bloom management.

Intermittent disturbance combined N and P adding favor colony size and abundance of Microcystis flos-aquae

Nutrients and disturbance are both important environmental factors in the freshwater ecosystem. Here we present data on the effects of intermittent disturbance (ID), nitrogen and phosphorus adding (+N+P), and intermittent disturbance combined N and P adding (ID+N+P) on colony size and abundance of Microcystis flos-aquae in lab experiments. Results showed that the mean colony sizes of M. flos-aquae in ID+N+P group were1.68, 1.56, 1.17 times that in the control, +N+P, ID groups, respectively. Moreover, the average number of M. flos-aquae in ID+N+P group were 1.47, 1.19, 1.42 times those in the control, +N+P, ID groups, respectively. The average concentration of EPS (extracellular polysaccharides) in ID and ID+N+P group was significantly higher than those in control and +N+P groups. Results demonstrated that intermittent disturbance combined N and P adding promoted colony size and abundance of M. flos-aquae. These results suggest that intermittent disturbance combined N and P adding plays an important role in the formation of Microcystis blooms in freshwater ecosystems.

Intermittent disturbance benefits colony size, biomass and dominance of Microcystis in Lake Taihu under field simulation condition

Wind-driven wave disturbance is one of the environmental factors that shapes the formation of Microcystis blooms. Here we present data on the effect of different disturbance modes (continuous vs intermittent disturbances) on colony size, biomass and dominance of Microcystis in Lake Taihu under field conditions. Small submersible pumps were used to simulate different disturbance modes at turbulent dissipation rate (ε) of 2.98 × 10−6 m2 s–3. Our results show that the mean colony sizes of Microcystis in intermittent and continuous disturbance group were 1.94 and 1.23 times that of the control group, respectively. The mean densities of Microcystis in intermittent and continuous disturbance group were 4.23 and 2.91 times that of the control group, respectively. The mean proportion of Microcystis to total algae abundance in control group and continuous disturbance group changed from 78.3% at beginning of the experiment to 4.5% and 9.1% at the end of the experiment. However, the proportion of Microcystis to total algae cells in intermittent disturbance group was 65.7–94.3% during the whole experiment. The results demonstrated intermittent disturbances favored colony morphology, biomass and dominance of Microcystis. Our results suggested that intermittent disturbance benefited the formation of Microcystis bloom and was important in the development of predictive models for toxic cyanobacterial blooms under changing climates in shallow lakes.

Cyanobacterial Dihydroxyacid Dehydratases Are a Promising Growth Inhibition Target.

Microbes are essential to the global ecosystem, but undesirable microbial growth causes issues ranging from food spoilage and infectious diseases to harmful cyanobacterial blooms. The use of chemicals to control microbial growth has achieved significant success, while specific roles for a majority of essential genes in growth control remain unexplored. Here, we show the growth inhibition of cyanobacterial species by targeting an essential enzyme for the biosynthesis of branched-chain amino acids. Specifically, we report the biochemical, genetic, and structural characterization of dihydroxyacid dehydratase from the model cyanobacterium Synechocystis sp. PCC 6803 (SnDHAD). Our studies suggest that SnDHAD is an oxygen-stable enzyme containing a [2Fe-2S] cluster. Furthermore, we demonstrate that SnDHAD is selectively inhibited in vitro and in vivo by the natural product aspterric acid, which also inhibits the growth of representative bloom-forming Microcystis and Anabaena strains but has minimal effects on microbial pathogens with [4Fe-4S] containing DHADs. This study suggests DHADs as a promising target for the precise growth control of microbes and highlights the exploration of other untargeted essential genes for microbial management.

Development of thermal treatment for the extraction of extracellular polymeric substances from Microcystis: Evaluating extraction efficiency and cell integrity

Extracellular polymeric substances (EPS) play a crucial role in processes associated with the growth and survival of microorganisms, such as in colony formation of cyanobacterium Microcystis. Thermal treatment is an effective method for EPS extraction. However, intracellular materials, however, often contaminate EPS extracts, possibly owing to cell lysis during thermal extraction. In this study, EPS samples were extracted using boric acid buffer under different temperatures (45–75 °C) and extraction periods (15–30 min), and evaluated with respect to both extraction efficiency and cell integrity. Moreover, the method developed in this study was compared to two widely employed methods (thermal extraction with 0.05% NaCl or NaOH (pH = 10)) with respect to EPS yield and composition, cell integrity or the release of cellular inclusions, and changes in cell size and morphology. Results indicated that thermal extraction with the boric acid buffer achieved the highest EPS yield and the lowest cell lysis. The optimal results were achieved by treatment at 65 °C for 30 min. However, a large quantity of cell debris and released cellular inclusions were detected when the other thermal methods were used, which suggests that stabilizing the cellular osmotic pressure and pH can effectively improve EPS yield and minimize cell lysis during thermal extraction. The findings of this study improve the EPS extraction from Microcystis, which can foster further investigations on the functions of these substances in the formation of Microcystis blooms.

Spatiotemporal dynamics of cell abundance, colony size and intracellular toxin concentrations of pelagic and benthic Microcystis in Lake Caohai, China

Lake Caohai has experienced extensive Microcystis blooms in recent years, and to improve its water quality, the local government carried out a series of water control measures. To better understand the dynamics of both pelagic and benthic Microcystis and their characteristics in Lake Caohai, we conducted a 1-year investigation from December 2015 to December 2016 to gain a seasonal outlook on the distribution and dynamics of cell abundance, colony size and intracellular microcystins (MCs) of Microcystis. The results indicated that the Microcystis bloom occupied primarily the northeastern region and then moved gradually from lakeshore to lake center. The perennial southwesterly winds and the water inflow from northeast to southwest in Lake Caohai determined the spatiotemporal distribution of pelagic Microcystis. Benthic Microcystis was mainly distributed in the northeastern region in summer, occupied the lake center in autumn and then occupied the southeastern region in winter, determined by the sedimentation of pelagic Microcystis and the death of benthic Microcystis. Small colonies (20–60 μm) overwintered more easily in both water column and sediment. The concentrations of intracellular toxin of benthic Microcystis were observed to be significantly higher than those of pelagic Microcystis. This might be because Microcystis synthesized large amount of MCs to acclimate to an unfavorable benthic environment. This knowledge on the dynamics of Microcystis expands our understanding of mechanisms underpinning the formation of Microcystis blooms.

Effects of secondary metabolites produced by different cyanobacterial populations on the freshwater zooplankters Brachionus calyciflorus and Daphnia pulex

Cyanobacterial blooms in eutrophic water bodies are a worldwide problem. Combined effects of mixtures of secondary metabolites produced by different cyanobacterial species on aquatic fauna are still not well recognised. We compared the survivorship of Brachionus calyciflorus Pallas (Rotifera) and Daphnia pulex Leyding (Cladocera) exposed to pure microcystin LR (MC-LR), anatoxin-a (ANTX) and to five extracts obtained from bloom-forming cyanobacteria Microcystis, Planktothrix and Dolichospermum. The obtained results revealed different response of the organisms to high concentrations of pure MC-LR, ANTX and complex cyanobacterial extracts. The extracts’ toxicity to invertebrates was higher than that exerted by pure cyanotoxins and was dependent on the composition of cyanobacterial metabolites: Microcystis spp. extract containing anabaenopeptins A and B, aeruginosamide, four variants of cyanopeptolins and five MCs was not toxic to either of the organisms, whereas Planktothrix agardhii extract (I), containing anabaenopeptins A, B, F, 915, oscillamide Y, five different aeruginosins and four variants of MC was more toxic to daphnids than to rotifers. The extracts of another P. agarhdii (II) biomass and two different biomass samples of Dolichospermum spp. also affected survivorship of the rotifer and cladoceran, however, to various extent. It strongly suggests that non-ribosomal oligopeptides, other than MCs, had essential contribution to the observed toxicity to invertebrates and their effects on particular species or populations can vary depending on the secondary metabolite profiles of cyanobacteria.

Evaluation of Size Structure in Freshwater Cyanobacterial Populations: Methods to Quantify Risk Associated with Changes in Biomass and Microcystin Concentrations

Cyanobacterial populations in surface waters, including drinking water supplies and recreational waters, represent an ever present challenge for resource managers. As communities continuously respond to external and internal processes, dynamic profiles of composition, dominance, growth and toxigenicity emerge. In this study measures of size structure and biomass, quantified using light microscopy and fluorometry, were used to estimate microcystin concentrations through linear regression analysis. Toxigenic profiles using cyanobacterial biomass were developed for lakes dominated by Microcystis spp. and Dolichospermum spp., influenced by both genus-specific pigment concentrations as well as microcystin concentrations. Community composition (Log %MIC) and biomass were used to describe microcystin concentrations in mixed assemblages, where composition was the first input variable. The accessory photopigment phycocyanin was used to describe the linear relationship between the daily growth and net microcystin production rates in the bloom-forming Microcystis spp. samples, suggesting that this size-fractionated sample may provide indications of potential toxigenicity in the whole lake water sample. Future investigations using fluorometric evaluation of cyanobacterial populations could provide additional applications and metrics for use by resource managers to quantify risk association with elevated cyanotoxin concentrations.

Microbial Diversity and Toxin Risk in Tropical Freshwater Reservoirs of Cape Verde.

The Cape Verde islands are part of the African Sahelian arid belt that possesses an erratic rain pattern prompting the need for water reservoirs, which are now critical for the country’s sustainability. Worldwide, freshwater cyanobacterial blooms are increasing in frequency due to global climate change and the eutrophication of water bodies, particularly in reservoirs. To date, there have been no risk assessments of cyanobacterial toxin production in these man-made structures. We evaluated this potential risk using 16S rRNA gene amplicon sequencing and full metagenome sequencing in freshwater reservoirs of Cape Verde. Our analysis revealed the presence of several potentially toxic cyanobacterial genera in all sampled reservoirs. Faveta potentially toxic and bloom-forming Microcystis sp., dominated our samples, while a Cryptomonas green algae and Gammaproteobacteria dominated Saquinho and Poilão reservoirs. We reconstructed and assembled the Microcystis genome, extracted from the metagenome of bulk DNA from Faveta water. Phylogenetic analysis of Microcystis cf. aeruginosa CV01’s genome revealed its close relationship with other Microcystis genomes, as well as clustering with other continental African strains, suggesting geographical coherency. In addition, it revealed several clusters of known toxin-producing genes. This survey reinforces the need to better understand the country’s microbial ecology as a whole of water reservoirs on the rise.

The highly heterogeneous methylated genomes and diverse restriction-modification systems of bloom-forming Microcystis

The occurrence of harmful Microcystis blooms is increasing in frequency in a myriad of freshwater ecosystems. Despite considerable research pertaining to the cause and nature of these blooms, the molecular mechanisms behind the cosmopolitan distribution and phenotypic diversity in Microcystis are still unclear. We compared the patterns and extent of DNA methylation in three strains of Microcystis, PCC 7806SL, NIES-2549 and FACHB-1757, using Single Molecule Real-Time (SMRT) sequencing technology. Intact restriction-modification (R-M) systems were identified from the genomes of these strains, and from two previously sequenced strains of Microcystis, NIES-843 and TAIHU98. A large number of methylation motifs and R-M genes were identified in these strains, which differ substantially among different strains. Of the 35 motifs identified, eighteen had not previously been reported. Strain NIES-843 contains a larger number of total putative methyltransferase genes than have been reported previously from any bacterial genome. Genomic comparisons reveal that methyltransferases (some partial) may have been acquired from the environment through horizontal gene transfer.