Microplastics enhanced the allelopathy of pyrogallol on toxic Microcystis with additional risks: Microcystins release and greenhouse gases emissions.

Cyanobacteria blooms (CBs) caused by eutrophication pose a global concern, especially Microcystis aeruginosa (M. aeruginosa), which could release harmful microcystins (MCs). The impact of microplastics (MPs) on allelopathy in freshwater environments is not well understood. This study examined the joint effect of adding polystyrene (PS-MPs) as representative MPs and two concentrations (2 and 8 mg/L) of pyrogallol (PYR) on the allelopathy of M. aeruginosa. The results showed that the addition of PS-MPs intensified the inhibitory effect of 8 mg/L PYR on the growth and photosynthesis of M. aeruginosa. After a 7-day incubation period, the cell density decreased to 69.7 %, and the chl-a content decreased to 48 % compared to the condition without PS-MPs (p < 0.05). Although the growth and photosynthesis of toxic Microcystis decreased with the addition of PS-MPs, the addition of PS-MPs significantly resulted in a 3.49-fold increase in intracellular MCs and a 1.10-fold increase in extracellular MCs (p < 0.05). Additionally, the emission rates of greenhouse gases (GHGs) (carbon dioxide, nitrous oxide and methane) increased by 2.66, 2.23 and 2.17-fold, respectively (p < 0.05). In addition, transcriptomic analysis showed that the addition of PS-MPs led to the dysregulation of gene expression related to DNA synthesis, membrane function, enzyme activity, stimulus detection, MCs release and GHGs emissions in M. aeruginosa. PYR and PS-MPs triggered ROS-induced membrane damage and disrupted photosynthesis in algae, leading to increased MCs and GHG emissions. PS-MPs accumulation exacerbated this issue by impeding light absorption and membrane function, further heightening the release of MCs and GHGs emissions. Therefore, PS-MPs exhibited a synergistic effect with PYR in inhibiting the growth and photosynthesis of M. aeruginosa, resulting in additional risks such as MCs release and GHGs emissions. These results provide valuable insights for the ecological risk assessment and control of algae bloom in freshwater ecosystems.

Can a shift in dominant species of Microcystis alter growth and reproduction of waterfleas?

The bloom-forming species Microcystis wesenbergii and M. aeruginosa occur in many lakes globally, and may exhibit alternating blooms both spatially and temporally. As environmental changes increase, cyanobacteria bloom in more and more lakes and are often dominated by M. wesenbergii. The adverse impact of M. aeruginosa on co-existing organisms including zooplanktonic species has been well-studied, whereas studies of M. wesenbergii are limited. To compare effects of these two species on zooplankton, we explored effects of exudates from different strains of microcystin-producing M. aeruginosa (Ma905 and Ma526) and non-microcystin-producing M. wesenbergii (Mw908 and Mw929), on reproduction by the model zooplankter Daphnia magna in both chronic and acute exposure experiments. Specifically, we tested physiological, biochemical, molecular and transcriptomic characteristics of D. magna exposed to Microcystis exudates. We observed that body length and egg and offspring number of the daphnid increased in all treatments. Among the four strains tested, Ma526 enhanced the size of the first brood, as well as total egg and offspring number. Microcystis exudates stimulated expression of specific genes that induced ecdysone, juvenile hormone, triacylglycerol and vitellogenin biosynthesis, which, in turn, enhanced egg and offspring production of D. magna. Even though all strains of Microcystis affected growth and reproduction, large numbers of downregulated genes involving many essential pathways indicated that the Ma905 strain might contemporaneously induce damage in D. magna. Our study highlights the necessity of including M. wesenbergii into the ecological risk evaluation of cyanobacteria blooms, and emphasizes that consequences to zooplankton may not be clear-cut when assessments are based upon production of microcystins alone.

Monitoring of toxic cyanobacterial blooms in Lalla Takerkoust reservoir by satellite imagery and microcystin transfer to surrounding farms.

Cyanobacterial harmful algal blooms (CyanoHABs) threaten public health and freshwater ecosystems worldwide. In this study, our main goal was to explore the dynamics of cyanobacterial blooms and how microcystins (MCs) move from the Lalla Takerkoust reservoir to the nearby farms. We used Landsat imagery, molecular analysis, collecting and analyzing physicochemical data, and assessing toxins using HPLC. Our investigation identified two cyanobacterial species responsible for the blooms: Microcystis sp. and Synechococcus sp. Our Microcystis strain produced three MC variants (MC-RR, MC-YR, and MC-LR), with MC-RR exhibiting the highest concentrations in dissolved and intracellular toxins. In contrast, our Synechococcus strain did not produce any detectable toxins. To validate our Normalized Difference Vegetation Index (NDVI) results, we utilized limnological data, including algal cell counts, and quantified MCs in freeze-dried Microcystis bloom samples collected from the reservoir. Our study revealed patterns and trends in cyanobacterial proliferation in the reservoir over 30 years and presented a historical map of the area of cyanobacterial infestation using the NDVI method. The study found that MC-LR accumulates near the water surface due to the buoyancy of Microcystis. The maximum concentration of MC-LR in the reservoir water was 160 µg L-1. In contrast, 4 km downstream of the reservoir, the concentration decreased by a factor of 5.39 to 29.63 µgL-1, indicating a decrease in MC-LR concentration with increasing distance from the bloom source. Similarly, the MC-YR concentration decreased by a factor of 2.98 for the same distance. Interestingly, the MC distribution varied with depth, with MC-LR dominating at the water surface and MC-YR at the reservoir outlet at a water depth of 10 m. Our findings highlight the impact of nutrient concentrations, environmental factors, and transfer processes on bloom dynamics and MC distribution. We emphasize the need for effective management strategies to minimize toxin transfer and ensure public health and safety.

A non-microcystin-producing Microcystis wesenbergii strain alters fish food intake by disturbing neuro-endocrine appetite regulation.

Cyanobacterial harmful algal blooms (cHABs) are pervasive sources of stress resulting in neurotoxicity in fish. A member of the widely distributed Microcystis genus of bloom-forming cyanobacteria, Microcystis wesenbergii can be found in many freshwater lakes, including Dianchi Lake (China), where it has become one of the dominant contributors to the lake's recurrent blooms. However, unlike its more well-known counterpart M. aeruginosa, the effects of dense non-microcystin-containing M. wesenbergii blooms are seldom studied. The disturbance of appetite regulation and feeding behaviour can have downstream effects on the growth of teleost fish, posing a significant challenge to aquaculture and conservation efforts. Here we examined the effects of M. wesenbergii blooms on the food intake of Acrossocheilus yunnanensis, a native cyprinid in southern China. This fish species has disappeared in Dianchi Lake, and its reintroduction might be negatively affected by the presence of this newly-dominant Microcystis species. We co-cultured juvenile A. yunnanensis with a non-microcystin-producing strain of M. wesenbergii at initial densities between 5 × 104 and 1 × 106 cells/mL and monitored fish feeding behaviour and changes in neurotransmitter and hormone protein levels. High-density M. wesenbergii cultures increased the feeding rate of co-cultured fish, elevating concentrations of appetite-stimulating signalling molecules (Agouti-related protein and γ-aminobutyric acid), while decreasing inhibitory ones (POMC). These changes coincided with histopathological alterations and reduced somatic indices in brain and intestinal tissues. Given this potential for detrimental effects and dysregulation of food intake, further studies are necessary to determine the impacts of chronic exposure of M. wesenbergii in wild fish.

Algae blooms with resistance in fresh water: Potential interplay between Microcystis and antibiotic resistance genes.

Microcystis, a type of cyanobacteria known for producing microcystins (MCs), is experiencing a global increase in blooms. They have been recently recognized as potential contributors to the widespread of antibiotic resistance genes (ARGs). By reviewing approximately 150 pieces of recent studies, a hypothesis has been formulated suggesting that significant fluctuations in MCs concentrations and microbial community structure during Microcystis blooms could influence the dynamics of waterborne ARGs. Among all MCs, microcystin-LR (MC-LR) is the most widely distributed worldwide, notably abundant in reservoirs during summer. MCs inhibit protein phosphatases or increase reactive oxygen species (ROS), inducing oxidative stresses, enhancing membrane permeability, and causing DNA damage. This further enhances selective pressures and horizontal gene transfer (HGT) chances of ARGs. The mechanisms by which Microcystis regulates ARG dissemination have been systematically organized for the first time, focusing on the secretion of MCs and the alterations of bacterial community structure. However, several knowledge gaps remain, particularly concerning how MCs interfere with the electron transport chain and how Microcystis facilitates HGT of ARGs. Concurrently, the predominance of Microcystis forming the algal microbial aggregates is considered a hotspot for preserving and transferring ARGs. Yet, Microcystis can deplete the nutrients from other taxa within these aggregates, thereby reducing the density of ARG-carrying bacteria. Therefore, further studies are needed to explore the 'symbiotic - competitive' relationships between Microcystis and ARG-hosting bacteria under varied nutrient conditions. Addressing these knowledge gaps is crucial to understand the impacts of the algal aggregates on dynamics of waterborne antibiotic resistome, and underscores the need for effective control of Microcystis to curb the spread of antibiotic resistance. Constructed wetlands and photocatalysis represent advantageous strategies for halting the spread of ARGs from the perspective of Microcystis blooms, as they can effectively control Microcystis and MCs while maintaining the stability of aquatic ecosystem.

Spatio-temporal connectivity of a toxic cyanobacterial community and its associated microbiome along a freshwater-marine continuum.

Due to climate changes and eutrophication, blooms of predominantly toxic freshwater cyanobacteria are intensifying and are likely to colonize estuaries, thus impacting benthic organisms and shellfish farming representing a major ecological, health and economic risk. In the natural environment, Microcystis form large mucilaginous colonies that influence the development of both cyanobacterial and embedded bacterial communities. However, little is known about the fate of natural colonies of Microcystis by salinity increase. In this study, we monitored the fate of a Microcystis dominated bloom and its microbiome along a French freshwater-marine gradient at different phases of a bloom. We demonstrated changes in the cyanobacterial genotypic composition, in the production of specific metabolites (toxins and compatible solutes) and in the heterotrophic bacteria structure in response to the salinity increase. In particular M. aeruginosa and M. wesenbergii survived salinities up to 20. Based on microcystin gene abundance, the cyanobacteria became more toxic during their estuarine transfer but with no selection of specific microcystin variants. An increase in compatible solutes occurred along the continuum with extensive trehalose and betaine accumulations. Salinity structured most the heterotrophic bacteria community, with an increased in the richness and diversity along the continuum. A core microbiome in the mucilage-associated attached fraction was highly abundant suggesting a strong interaction between Microcystis and its microbiome and a likely protecting role of the mucilage against an osmotic shock. These results underline the need to better determine the interactions between the Microcystis colonies and their microbiome as a likely key to their widespread success and adaptation to various environmental conditions.

Potential gap in understanding cyanoHABs: Light-dependent morphological variations in colonial cyanobacterium Microcystis.

Colony formation is a crucial characteristic of Microcystis, a cyanobacterium known for causing cyanobacterial harmful algal blooms (cyanoHABs). It has been observed that as Microcystis colonies grow larger, they often become less densely packed, which correlates with a decrease in light penetration. The objective of this study was to investigate the effects of light limitation on the morphological variations in Microcystis, particularly in relation to the crowded cellular environment. The results indicated that when there was sufficient light (transmittance = 100 %) to support a growth rate of 0.11±0.01 day-1, a significant increase in colony size was found, from 466±15 µm to 1030±111 µm. However, under light limitation (transmittance = 50 % - 1 %) where the growth rate was lower than 0, there was no significant improvement in colony size. Microcystis in the light limitation groups exhibited a loose cell arrangement and even the presence of holes or pores within the colony, confirming the negative correlation between colony size and cell arrangement. This pattern is driven by regional differences in growth within the colony, as internal cells have a significantly lower frequency of division compared to peripheral cells, due to intra-colony self-shading (ICSS). The research demonstrates that Microcystis can adjust its cell arrangement to avoid excessive self-shading, which has implications for predicting and controlling cyanoHABs. These findings also contribute to the understanding of cyanobacterial variations and can potentially inform future research on the diverse phycosphere.

Combined effect of freshwater salinization and harmful algae on the benthic invertebrate Chironomus pallidivittatus.

Global climate change as well as human activities have been reported to increase the frequency and severity of both salinization and harmful algal blooms (HABs) in many freshwater systems, but their co-effect on benthic invertebrates has rarely been studied. This study simultaneously examined the joint toxicity of salinity and different cyanobacterial diets on the behavior, development, select biomarkers, and partial life cycle of Chironomus pallidivittatus (Diptera). High concentrations of salts (e.g., 1 g/L Ca2+ and Mg2+) and toxic Microcystis had synergistic toxicity, inhibiting development, burrowing ability and causing high mortality of C. pallidivittatus, especially for the Mg2+ treatment, which caused around 90% death. Low Ca2+ concentration (e.g., 0.01 g/L) promoted larval burrowing ability and inhibited toxin accumulation, which increased the tolerance of Chironomus to toxic Microcystis. However, low Mg2+ concentration (e.g., 0.01 g/L) was shown to inhibit the behavior, development and increase algal toxicity to Chironomus. Toxic Microcystis resulted in microcystin (MC) accumulation, inhibited the burrowing ability of larvae, and increased the proportion of male adults (>50%). The combined toxicity level from low to high was verified by the weight of evidence and the grey TOPSIS model, which integrated five lines of evidence to increase the risk assessment accuracy and efficiency. This is the first study that provided insights into ecological risk arising from the joint effect of salinity and harmful algae on benthic organisms. We suggest that freshwater salinization and HABs should be considered together when assessing ecological threats that arise from external stress.

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.

Mechanisms of harmful effects of Microcystis aeruginosa on a brackish water organism Moina mongolica based on physiological and transcriptomic responses.

To investigate the detrimental impacts of cyanobacterial bloom, specifically Microcystis aeruginosa, on brackish water ecosystems, the study used Moina mongolica, a cladoceran species, as the test organism. In a chronic toxicology experiment, the survival and reproductive rates of M. mongolica were assessed under M. aeruginosa stress. It was observed that the survival rate of M. mongolica fed with M. aeruginosa significantly decreased with time and their reproduction rate dropped to zero, while the control group remained maintained stable and normal reproduction. To further explore the underlying molecular mechanisms of the effects of M. aeruginosa on M. mongolica, we conducted a transcriptomic analysis on newly hatched M. mongolica cultured under different food conditions for 24 h. The results revealed significant expression differences in 572 genes, with 233 genes significantly up-regulated and 339 genes significantly down-regulated. Functional analysis of these differentially expressed genes identified six categories of physiological functional changes, including nutrition and metabolism, oxidative phosphorylation, neuroimmunology, cuticle and molting, reproduction, and programmed cell death. Based on these findings, we outlined the basic mechanisms of microcystin toxicity. The discovery provides critical insights into the mechanisms of Microcystis toxicity on organisms and explores the response mechanisms of cladocerans under the stress of Microcystis.

Selection of photosynthetic traits by turbulent mixing governs formation of cyanobacterial blooms in shallow eutrophic lakes.

Prediction of the complex cyanobacteria-environment interactions is vital for understanding harmful bloom formation. Most previous studies on these interactions considered specific properties of cyanobacterial cells as representative for the entire population (e.g. growth rate, mortality, and photosynthetic capacity (Pmax)), and assumed that they remained spatiotemporally unchanged. Although, at the population level, the alteration of such traits can be driven by intraspecific competition, little is known about how traits and their plasticity change in response to environmental conditions and affect the bloom formation. Here we test the hypothesis that intraspecific variations in Pmax of cyanobacteria (Microcystis spp.) play an important role in its population dynamics. We coupled a one-dimensional hydrodynamic model with a trait-based phytoplankton model to simulate the effects of physical drivers (turbulence and turbidity) on the Pmax of Microcystis populations for a range of dynamic conditions typical for shallow eutrophic lakes. Our results revealed that turbulence acts as a directional selective driver for changes in Pmax. Depending on the intensity of daily-periodic turbulence, representing wind-driven mixing, a shift in population-averaged phenotypes occurred toward either low Pmax, allowing the population to capture additional light in the upper layers, or high Pmax, enhancing the efficiency of light utilization. Moreover, we observed that a high intraspecific diversity in Pmax accelerated the formation of surface scum by up to more than four times compared to a lower diversity. This study offers insights into mechanisms by which cyanobacteria populations respond to turbulence and underscores the significance of intraspecific variations in cyanobacterial bloom formation.

Upstream nitrogen availability determines the Microcystis salt tolerance and influences microcystins release in brackish water.

The occurrence of large Microcystis biomass in brackish waters is primarily caused by its downward transportation from the upstream freshwater lakes and reservoirs through rivers rather than due to in situ bloom formation. Factors that determine the survival of freshwater cyanobacteria in brackish waters have not been well investigated. Here, we studied the spatiotemporal variability of inorganic nitrogen in an upstream lake and conducted laboratory and in-situ experiments to assess the role of nitrogen availability on the salt tolerance of Microcystis and the release of microcystins. A series of field experiments were carried out during bloom seasons to evaluate the salt tolerance of natural Microcystis colonies. The salt tolerance threshold varied from 7 to 17 and showed a positive relationship with intracellular carbohydrate content and a negative relationship with nitrogen availability in water. In August when upstream nitrogen availability was lower, the Microcystis colonies could maintain their biomass even after a sudden increase in salinity from 4 to 10. Laboratory-cultivated Microcystis that accumulated higher carbohydrate content at lower nitrogen availability showed better cell survival at higher salinity. The sharp release of microcystins into the surrounding water occurred when salinity exceeded the salt tolerance threshold of the Microcystis. Thus, Microcystis with higher salt tolerance can accumulate more toxins in cells. The obtained results suggest that the cell survival and toxin concentration in brackish waters depend on the physiological properties of Microcystis formed in the upstream waters. Thus, the life history of Microcystis in upstream waters could have a significant impact on its salt tolerance in downstream brackish waters, where the ecological risk of the salt-tolerant Microcystis requires special and careful management in summer at low nitrogen availability.

Combined effects of spinetoram and Microcystis aeruginosa on Daphnia pulex offspring: Maternal effects and multigenerational implications.

The increasing occurrence of harmful algal blooms (HABs) in freshwater ecosystems detrimentally affect global water environments. Zooplankton's role in controlling HABs is hindered by contaminant exposure, necessitating research into combined stressors' ecological impacts. The response of Daphnia, a freshwater keystone species, to environmental stressors can be influenced by its maternal effects. Here, we investigated the combined effects of the world-widely used insecticide spinetoram and non-toxic HABs species Microcystis aeruginosa on the life-history traits of D. pulex offspring produced from different maternal food conditions. Four maternal groups were established, with each group receiving a specific blend of C. vulgaris (Ch) and M. aeruginosa (Ma) in varying proportions: A (100% Ch), B (90% Ch + 10% Ma), C (80% Ch + 20% Ma), and D (70% Ch + 30% Ma). The offspring from the third brood were gathered, and a 21-day experiment was carried out, involving various feeding groups (AA, AD, BA, BB, CA, CC, DA, and DD). Results demonstrated that grazing on M. aeruginosa by D. pulex induced maternal effects on their offspring, with the continuous exposure group showing an enhanced tolerance to M. aeruginosa. This study also unveiled that spinetoram could interfere with the molting of D. pulex, leading to developmental retardation. The Recovery Group exhibited an intriguing phenomenon: under the influence of both concentrations of the pesticide spinetoram (0.18, 0.35 µg L-1), D. pulex produced more offspring. This might be due to a combined strategy of allocating more energy towards reproduction in response to low-quality food and a potential hormetic effect from low concentrations of spinetoram. Assessing the interplay of combined stressors across multiple generations, encompassing harmful algal blooms (HABs) and environmental pollutants, is essential for predicting population responses to evolving environmental conditions. This understanding is vital for the protection and management of aquatic environments and ecosystems.

Physiology, microcystin production, and transcriptomic responses of Microcystis aeruginosa exposed to calcium and magnesium.

Calcium ions (Ca2+) and magnesium ions (Mg2+) are pivotal in the community composition and stability of harmful cyanobacteria, yet the physiological and molecular responses remains poorly understood. This study aims to explore these responses in the high microcystin producer Microcystis aeruginosa (M. aeruginosa). Results indicate that the growth of M. aeruginosa is inhibited by Ca2+/Mg2+ exposure (0.5-10 mM), while Fv/Fm photosynthetic parameters and extracellular microcystin-leucine-arginine (MC-LR) concentrations increase. Additionally, MC-LR release is significantly elevated under exposure to Ca2+/Mg2+, posing potential aquatic environmental risks. Transcriptomic analysis reveals downregulation of genes related to cell architecture, membrane transport, and metabolism, while the genes linked to photosynthesis electron transmission and heavy metal-responsive transcriptional regulators are upregulated to adapt to environmental changes. Further analysis reveals that Ca2+ and Mg2+ primarily impact sulfur metabolism and transport of amino acids and mineral within cells. These findings provide insights into M. aeruginosa cells responses to Ca2+ and Mg2+ exposure.

The toxicological effects of life-cycle exposure to harmful benthic cyanobacteria Oscillatoria on zebrafish growth and reproduction: A comparative study with planktonic Microcystis.

The risks of planktonic cyanobacteria blooms have been the focus of much scientific research, but studies on the ecotoxicological effects of benthic cyanobacteria are lagging. The impacts of cyanobacteria cells on fish populations might be more complex in contrast to purified cyanotoxins or cyanobacteria extracts. This study systematically compared the chronic effects of benthic Oscillatoria sp. (producing cylindrospermopsins) and planktonic Microcystis aeruginosa (producing microcystins) on the growth and reproduction of zebrafish through life-cycle exposure (5- 90 days post fertilization). The results showed that both Oscillatoria sp. and M. aeruginosa exposure caused growth inhibition and fecundity reduction in F0 generation by disrupting sex hormone levels, delayed ovarian and sperm development, and induced pathological lesions in zebrafish gonads. Furthermore, exposure to Oscillatoria sp. or M. aeruginosa in adult zebrafish increased mortality and teratogenicity in F1 embryos (without exposure), indicating a parental transmission effect of developmental toxicity. The difference was that M. aeruginosa exposure led to significant alterations in pathways, such as tissue development, redox processes, and steroid hormone synthesis. In contrast, Oscillatoria sp. exposure primarily disrupted the PPAR signaling pathway, cell adhesion molecules, and lipid transport pathways. Interestingly, the differentially expressed genes revealed that male fish were more sensitive to harmful cyanobacteria than females, whether exposed to Oscillatoria sp. or M. aeruginosa. These findings contribute to a better mechanistic understanding of the chronic toxic effects of distinct types of harmful cyanobacteria, suggesting that the ecological risk of benthic cyanobacteria requires further attention.

Elimination of Microcystis aeruginosa through Leuconostoc mesenteroides DH and its underlying mechanism.

Microcystis aeruginosa is ubiquitously found in various water bodies and can produce microcystins (MCs), which threaten the health of aquatic animals and human beings. The elimination of excessive M. aeruginosa is beneficial for the protection of the ecosystems and public health. In this regard, algae-lysing bacteria have been extensively studied as an effective measure for their eradication. However, the active substances generated by algae-lysing bacteria are limited. For this study, we reveal that the phenyllactic acid (PLA) produced by Leuconostoc mesenteroides DH exhibits high efficacy for the removal of M. aeruginosa, and explore the elimination mechanism of strain DH on M. aeruginosa. It was found that a cell-free supernatant of strain DH possessed high removal activities against M. aeruginosa. Abundant reactive oxygen species were induced in algal cells following exposure to strain DH supernatant, as well as superoxide dismutase and catalase responses. Furthermore, the integrity of algal cell membranes and photosynthesis was seriously damaged. Interestingly, added exogenous eugenol significantly inhibited the synthesis of active substance produced by strain DH, which further identified that PLA is one of the active substances that contribute to the eradication of M. aeruginosa on the basis of metabolomics analysis. Our finding demonstrated, for the first time, that PLA (as an anti-cyanobacterial compound) can be used for the removal of M. aeruginosa, which provides a theoretical basis for the control of M. aeruginosa.

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.

Insights from colony formation: The necessity to consider morphotype when assessing the effect of antibiotics on cyanobacteria.

Colonial cyanobacteria have been identified as the primary contributor to the global occurrence of cyanobacterial harmful algal blooms (cyanoHABs), which are further intensified by the presence of "pseudo-persistent" antibiotics. Nevertheless, the impact of antibiotics on the growth and size of colonial cyanobacteria remains unclear. In this study, the response of cyanobacterium Microcystis to varying doses of antibiotics was assessed (0, 0.1, 0.5, 1, 10, and 50 µg L-1) by comparing the unicellular and colonial morphotypes. Interestingly, the morphological structure of cyanobacteria plays a significant role in their reaction to antibiotics. In comparison to the unicellular morphotype, the colonial morphotype exhibited a greater promotion in growth rate (11 %-22 %) to low doses of antibiotics and was less inhibited (-121 %--62 %) under high doses. Furthermore, antibiotics may affect the size of cyanobacterial colonies by disrupting the secretion of algal organic matter, which also exhibited a two-phase pattern. This work sheds light on the significance of methodology research involving both unicellular and colonial cyanobacteria. Future research and lake management should prioritize studying the morphological traits of cyanobacteria under different levels of antibiotic exposure. This approach may lead to novel strategies for predicting cyanoHABs under antibiotic pollution more effectively.

Exploring an Efficient and Eco-Friendly Signaling Molecule and Its Quorum Quenching Ability for Controlling Microcystis Blooms.

Globally, cyanobacterial blooms have become serious problems in eutrophic water. Most previous studies have focused on environmental factors but have neglected the role of quorum sensing (QS) in bloom development and control. This study explored a key quorum sensing molecule (QSM) that promotes cell growth and then proposed a targeted quorum quencher to control blooms. A new QSM 3-OH-C4-HSL was identified with high-resolution mass spectrometry. It was found to regulate cellular carbon metabolism and energy metabolism as a means to promote Microcystis aeruginosa growth. To quench the QS induced by 3-OH-C4-HSL, three furanone-like inhibitors were proposed based on molecular structure, of which dihydro-3-amino-2-(3H)-furanone (FN) at a concentration of 20 µM exhibited excellent inhibition of M. aeruginosa growth (by 67%). Molecular docking analysis revealed that the inhibitor strongly occupied the QSM receptor protein LuxR by binding with Asn164(A) and His167(A) via two hydrogen bonds (the bond lengths were 3.04 and 4.04 Å) and the binding energy was -5.9 kcal/mol. The inhibitor blocked signaling regulation and induced programmed cell death in Microcystis. Importantly, FN presented little aquatic biotoxicity and negligibly affected aquatic microbial function. This study provides a promising new and eco-friendly strategy for controlling cyanobacterial blooms.

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.