Changes of vertical distribution of Microcystis colonies driven by short-term rainfall: Disappearance and reformation of surface bloom.

Global climate warming and human activities have increased the magnitude and frequency of Microcystis surface blooms, posing significant threats to freshwater ecosystems and human health over recent decades. Heavy rainfall events have been reported to cause the disappearance of these blooms. Although some studies have employed turbulence models to analyze the movement characteristics of Microcystis colonies, the impact of rainfall is complex, comprehensive investigations on their vertical migration induced by short-term rainfall are still necessary. Utilizing monitoring data from eutrophic ponds and controlled simulation experiments, this study examines the short-term impacts of rainfall on the vertical distribution of Microcystis in the water column. Our findings indicate that rainfall contributes to the disappearance of Microcystis blooms by reducing the quantity of small to medium-sized colonies (0-100 µm) at the surface, subsequently decreasing the overall Microcystis biomass. As rainfall intensity increases, larger colonies migrate deeper into the water column. At a rainfall threshold of 666 mm, the difference in the median volume diameter (DV50) of Microcystis colonies between the surface and bottom reaches a minimal value of 3.09%. Post-rainfall, these colonies rapidly ascend, aggregate into larger formations, and re-establish surface blooms. The greater the rainfall, the smaller the resultant Microcystis biomass, albeit with larger aggregated colony sizes. When rainfall exceeds 222 mm, the recovery rate of surface Microcystis biomass remains below 100%, decreasing to 19.48% at 666 mm of rainfall, while the median volume diameter (DV50) of the colonies increases to 139.07% of its pre-rainfall level. Furthermore, compared to pre-rainfall conditions, the photosynthetic activity of the surface Microcystis colonies was enhanced and the secretion of EPS was increased under heavy rainfall conditions. Our results identify a critical response time of 30 min for Microcystis colonies to rainfall, after which the response ceases to intensify. These insights are crucial for predicting post-rain Microcystis bloom dynamics and aiding management authorities in timely interventions.

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.

Changes in CO2 concentration drive a succession of toxic and non-toxic strains of Microcystis blooms.

The dynamic changes between toxic and non-toxic strains of Microcystis blooms have always been a hot topic. Previous studies have found that low CO2 favors toxic strains, but how changing dissolved CO2 (CO2 [aq]) in water body influences the succession of toxic and non-toxic strains in Microcystis blooms remains uncertain. Here, we combined laboratory competition experiments, field observations, and a machine learning model to reveal the links between CO2 changes and the succession. Laboratory experiments showed that under low CO2 conditions (100-150 ppm), the toxic strains could make better use of CO2 (aq) and be dominant. The non-toxic strains demonstrated a growth advantage as CO2 concentration increased (400-1000 ppm). Field observations from June to November in Lake Taihu showed that the percentage of toxic strains increased as CO2 (aq) decreased. Machine learning highlighted links between the inorganic carbon concentration and the proportion of advantageous strains. Our findings provide new insights for cyanoHABs prediction and prevention.

Comparative study about ozonation to treat Microcystis-laden source water at the development and maintenance stage.

The outbreak of toxic cyanobacteria blooms is hazardous to water safety. Ozonation has been used to treat cyanobacteria-laden source water. Generally, cyanobacterial blooms enter into a long-term maintenance stage from the bloom development, but how the changed bloom stage affects ozonation is still unknow. Herein, influences of ozonation on cell inactivation and microcystin removal of Microcystis at the development and maintenance stage, were investigated. Then, ozonation-assisted coagulation for Microcystis removal at the two stages was compared. Results showed no significant difference in the photosynthetic inactivation of Microcystis at both stages. Microcystis at the maintenance stage exhibited a lower loss of membrane integrity (268-480 M-1 s-1) than that at the development stage (413-596 M-1 s-1). However, the extracellular microcystin increased by 30-410% at the maintenance stage at a lower ratio of [O3: DOC] (0.10-0.80) compared to the development stage (0.21-1.68), mainly ascribed to a decrease in the ozonation efficiency for microcystin removal. This finding might result from the elevated biomass and N-containing organics as competitors to reduce microcystin ozonation. Meanwhile, it was possible to generate fewer hydroxyl radicals to oxidize microcystin at the maintenance stage than that at the development stage. Besides, the removal ratio of Microcystis after ozonation-assisted coagulation, was reduced by 46-230% at the maintenance stage, due to the insufficient modification of cellular surface or elevated organics of 3-30 kDa. This work indicated that ozonation is effective to treat Microcystis at the development stage of a bloom whist pre-ozonation might be an inappropriate choice at the long-term maintenance stage.

The cyanobactericidal bacterium Paucibacter aquatile DH15 caused the decline of Microcystis and aquatic microbial community succession: A mesocosm study.

Microcystis blooms pose a major threat to the quality of drinking water. Cyanobactericidal bacteria have attracted much attention in the research community as a vehicle for controlling Microcystis blooms because of their ecological safety. Nonetheless, most studies on cyanobactericidal bacteria have been conducted on a laboratory scale but have not been scaled-up as field experiments. Thus, our understanding of the microbial response to cyanobactericidal bacteria in natural ecosystems remains elusive. Herein, we applied Paucibacter aquatile DH15 to control Microcystis blooms in a 1000 L mesocosm experiment and demonstrated its potential with the following results: (1) DH15 reduced Microcystis cell density by 90.7% within two days; (2) microcystins released by Microcystis death decreased to the control level in four days; (3) during the cyanobactericidal processes, the physicochemical parameters of water quality remained safe for other aquatic organisms; and (4) the cyanobactericidal processes promoted the growth of eukaryotic microalgae, replacing cyanobacteria. The cyanobactericidal processes accelerated turnover rates, decreased stability, and altered the functional profile of the microbial community. Network analysis demonstrated that this process resulted in more complex interactions between microbes. Overall, our findings suggest that strain DH15 could be considered a promising candidate for controlling Microcystis blooms in an eco-friendly manner.

Revealing Physiochemical Factors and Zooplankton Influencing Microcystis Bloom Toxicity in a Large-Shallow Lake Using Bayesian Machine Learning.

Toxic cyanobacterial blooms have become a severe global hazard to human and environmental health. Most studies have focused on the relationships between cyanobacterial composition and cyanotoxins production. Yet, little is known about the environmental conditions influencing the hazard of cyanotoxins. Here, we analysed a unique 22 sites dataset comprising monthly observations of water quality, cyanobacterial genera, zooplankton assemblages, and microcystins (MCs) quota and concentrations in a large-shallow lake. Missing values of MCs were imputed using a non-negative latent factor (NLF) analysis, and the results achieved a promising accuracy. Furthermore, we used the Bayesian additive regression tree (BART) to quantify how Microcystis bloom toxicity responds to relevant physicochemical characteristics and zooplankton assemblages. As expected, the BART model achieved better performance in Microcystis biomass and MCs concentration predictions than some comparative models, including random forest and multiple linear regression. The importance analysis via BART illustrated that the shade index was overall the best predictor of MCs concentrations, implying the predominant effects of light limitations on the MCs content of Microcystis. Variables of greatest significance to the toxicity of Microcystis also included pH and dissolved inorganic nitrogen. However, total phosphorus was found to be a strong predictor of the biomass of total Microcystis and toxic M. aeruginosa. Together with the partial dependence plot, results revealed the positive correlations between protozoa and Microcystis biomass. In contrast, copepods biomass may regulate the MC quota and concentrations. Overall, our observations arouse universal demands for machine-learning strategies to represent nonlinear relationships between harmful algal blooms and environmental covariates.

Prediction of Microcystis Occurrences and Analysis Using Machine Learning in High-Dimension, Low-Sample-Size and Imbalanced Water Quality Data.

Machine learning, Deep learning, and water quality data have been used in recent years to predict the outbreak of harmful algae, especially Microcystis, and analyze outbreak causes. However, for various reasons, water quality data are often High-Dimension, Low-Sample- Size (HDLSS), meaning the sample size is lower than the number of dimensions. Moreover, imbalance problems may arise due to bias in the occurrence frequency of Microcystis. These problems make predicting the occurrence of Microcystis and analyzing its causes with machine learning difficult. In this study, a machine learning model that applies Feature Engineering (FE) and Feature Selection (FS) algorithms are used to predict outbreaks of Microcystis and analyze the outbreak factors from imbalanced HDLSS water quality data. The prediction performance was verified with binary classification to determine whether Microcystis would occur in the future by applying three machine learning models to four data patterns. The cause analysis of Microcystis occurrence was performed by visualizing the results of applying FE and FS. For the test data, the predictive performance of FE and FS methods was significantly better than that of the conventional method, with an accuracy of .108 points and an F-value of .691 points higher than the conventional method. A prediction performance increase was observed with a smaller model capacity. Data-driven analysis suggested that total nitrogen, chemical oxygen demand, chlorophyll-a, dissolved oxygen saturation, and water temperature are associated with Microcystis occurrences. The results also indicated that basic statistics of the water quality distribution (especially mean, standard deviation, and skewness) over a year, not the concentrations of water components, are related to the occurrence of Microcystis. These are new findings not found in previous studies and are expected to contribute significantly to future studies of algae. This study provides a method for analyzing water quality data with high-dimensionality and small samples, imbalance problems, or both.

Gene expression pattern of microbes associated with large cyanobacterial colonies for a whole year in Lake Taihu.

Large cyanobacterial colonies, which are unique niches for heterotrophic bacteria, are vital for blooming in eutrophic waters. However, the seasonal dynamics of molecular insights into microbes in these colonies remain unclear. Here, the community composition and metabolism pattern of microbes inhabiting large cyanobacterial colonies (> 120 µm, collected from Lake Taihu in China) were investigated monthly. The community structure of total microbes was mostly influenced by chlorophyll a (Chl a), total phosphorus (TP) concentration, dissolved oxygen, and temperature, whereas the colony-associated bacteria (excluding Cyanobacteria) were mostly influenced by total organic carbon, NO3-, and PO43- concentrations, indicating different response patterns of Cyanobacteria and the associated bacteria to water nutrient conditions. Metatranscriptomic data suggested that similar to that of Cyanobacteria, the gene expression patterns of the most active bacteria, such as Proteobacteria and Bacteroidetes, were not strictly dependent on season but separated by Chl a concentrations. Samples in July and September (high-bloom period) and February and March (non-bloom period) formed two distinct clusters, whereas those of other months (low-bloom period) clustered together. The accumulation of transcripts for pathways, such as phycobilisome from Cyanobacteria and bacterial chemotaxis and flagellum, phosphate metabolism, and sulfur oxidation from Proteobacteria, was enriched in high- and low-bloom periods than in non-bloom period. Network analyses revealed that Cyanobacteria and Proteobacteria exhibited coordinated transcriptional patterns in almost all divided modules. Modules had Cyanobacteria-dominated hub gene were positively correlated with temperature, Chl a, total dissolved phosphorus, and NH4+ and NO2- concentrations, whereas modules had Proteobacteria-dominated hub gene were positively correlated with TP and PO43-. These results indicated labor division might exist in the colonies. This study provided metabolic insights into microbes in large cyanobacterial colonies and would support the understanding and management of the year-round cyanobacterial blooms.

Simultaneous Removal of the Freshwater Bloom-Forming Cyanobacterium Microcystis and Cyanotoxin Microcystins via Combined Use of Algicidal Bacterial Filtrate and the Microcystin-Degrading Enzymatic Agent, MlrA.

Freshwater cyanobacterial blooms (e.g., Microcystis blooms) constitute a major global environmental problem because of their risks to public health and aquatic ecological systems. Current physicochemical treatments of toxic cyanobacteria cause the significant release of cyanotoxin microcystins from damaged cells. Biological control is a promising eco-friendly technology to manage harmful cyanobacteria and cyanotoxins. Here, we demonstrated an efficient biological control strategy at the laboratory scale to simultaneously remove Microcystis and microcystins via the combined use of the algicidal bacterial filtrate and the microcystin-degrading enzymatic agent. The algicidal indigenous bacterium Paenibacillus sp. SJ-73 was isolated from the sediment of northern Lake Taihu, China, and the microcystin-degrading enzymatic agent (MlrA) was prepared via the heterologous expression of the mlrA gene in the indigenous microcystin-degrading bacterium Sphingopyxis sp. HW isolated from Lake Taihu. The single use of a fermentation filtrate (5%, v/v) of Paenibacillus sp. SJ-73 for seven days removed the unicellular Microcystis aeruginosa PCC 7806 and the native colonial Microcystis strain TH1701 in Lake Taihu by 84% and 92%, respectively, whereas the single use of MlrA removed 85% of microcystins. Used in combination, the fermentation filtrate and MlrA removed Microcystis TH1701 and microcystins by 92% and 79%, respectively. The present biological control thus provides an important technical basis for the further development of safe, efficient, and effective measures to manage Microcystis blooms and microcystins in natural waterbodies.

Elevated pH Conditions Associated With Microcystis spp. Blooms Decrease Viability of the Cultured Diatom Fragilaria crotonensis and Natural Diatoms in Lake Erie.

Cyanobacterial Harmful Algal Blooms (CyanoHABs) commonly increase water column pH to alkaline levels ≥9.2, and to as high as 11. This elevated pH has been suggested to confer a competitive advantage to cyanobacteria such as Microcystis aeruginosa. Yet, there is limited information regarding the restrictive effects bloom-induced pH levels may impose on this cyanobacterium's competitors. Due to the pH-dependency of biosilicification processes, diatoms (which seasonally both precede and proceed Microcystis blooms in many fresh waters) may be unable to synthesize frustules at these pH levels. We assessed the effects of pH on the ecologically relevant diatom Fragilaria crotonensis in vitro, and on a Lake Erie diatom community in situ. In vitro assays revealed F. crotonensis monocultures exhibited lower growth rates and abundances when cultivated at a starting pH of 9.2 in comparison to pH 7.7. The suppressed growth trends in F. crotonensis were exacerbated when co-cultured with M. aeruginosa at pH conditions and cell densities that simulated a cyanobacteria bloom. Estimates demonstrated a significant decrease in silica (Si) deposition at alkaline pH in both in vitro F. crotonensis cultures and in situ Lake Erie diatom assemblages, after as little as 48 h of alkaline pH-exposure. These observations indicate elevated pH negatively affected growth rate and diatom silica deposition; in total providing a competitive disadvantage for diatoms. Our observations demonstrate pH likely plays a significant role in bloom succession, creating a potential to prolong summer Microcystis blooms and constrain diatom fall resurgence.

The nitrogen reduction in eutrophic water column driven by Microcystis blooms.

During the bloom seasons, the dissolved inorganic nitrogen declines, which results in the occurrence of nitrogen limitation. It is unclear where the nitrogen goes. Our enclosure experiments and batch tests suggested that Microcystis blooms could significantly reduce the nitrogen in water bodies and the key mechanisms for the nitrogen reduction in different layers were different. The assimilation was the main pathway for nitrogen reduction in the surface layer, while denitrification played an important role both at the sediment-water interface and in the overlying water. Stable nitrogen isotope experiments showed that the nitrate reduction efficiency at sediment-water interface was enhanced by Microcystis, reaching to 76.5∼84.7 %. Dissimilation accounted for 63.8∼67.3 % of the nitrate reduction, and the denitrification rate was 7.4∼8.5 times of DNRA rate. In the water column, the Microcystis bloom facilitated the formation of dark/anoxic condition, which favored the denitrification. The Microcystis aggregates collected from the field showed a great potential in removing nitrogen, and the TN in the overly water was reduced by 3.76∼6.03 mg L-1 within two days. This study provided field evidences and deeper insights into the relationship between Microcystis blooms and nitrogen reduction in the whole water column and gave more details about the enhancing effects of Microcystis on nitrogen reduction.

How physiological and physical processes contribute to the phenology of cyanobacterial blooms in large shallow lakes: A new Euler-Lagrangian coupled model.

Cyanobacterial blooms have emerged as one of the most severe ecological problems affecting large and shallow freshwater lakes. To improve our understanding of the factors that influence, and could be used to predict, surface blooms, this study developed a novel Euler-Lagrangian coupled approach combining the Eulerian model with agent-based modelling (ABM). The approach was subsequently verified based on monitoring datasets and MODIS data in a large shallow lake (Lake Taihu, China). The Eulerian model solves the Eulerian variables and physiological parameters, whereas ABM generates the complete life cycle and transport processes of cyanobacterial colonies. This model ensemble performed well in fitting historical data and predicting the dynamics of cyanobacterial biomass, bloom distribution, and area. Based on the calculated physical and physiological characteristics of surface blooms, principal component analysis (PCA) captured the major processes influencing surface bloom formation at different stages (two bloom clusters). Early bloom outbreaks were influenced by physical processes (horizontal transport and vertical turbulence-induced mixing), whereas buoyancy-controlling strategies were essential for mature bloom outbreaks. Canonical correlation analysis (CCA) revealed the combined actions of multiple environment variables on different bloom clusters. The effects of buoyancy-controlling strategies (ISP), vertical turbulence-induced mixing velocity of colony (VMT) and horizontal drift velocity of colony (HDT) were quantitatively compared using scenario simulations in the coupled model. VMT accounted for 52.9% of bloom formations and maintained blooms over long periods, thus demonstrating the importance of wind-induced turbulence in shallow lakes. In comparison, HDT and buoyancy controlling strategies influenced blooms at different stages. In conclusion, the approach developed here presents a promising tool for understanding the processes of onshore/offshore algal blooms formation and subsequent predicting.

Colony formation in two Microcystis morphotypes: Effects of temperature and nutrient availability.

The ability of Microcystis to form large colonies is a key trait that contributes to competition ability over other phytoplankton and facilitates the formation of surface scums in many freshwater systems. The effect of temperature and nutrients on this trait, however, is far from clear and needs further investigation, especially under a warmer climate and nutrient overloading in aquatic systems globally. In this study, two colonial strains of Microcystis (M. wesenbergii and M. ichthyoblabe) originally isolated from Lake Taihu in China, were used to investigate cyanobacterial aggregation under a range of temperatures (15-30 °C), phosphorus availability (0.004-8 mg P L-1), and nitrogen availability (0.04-40 mg N L-1). The mechanism of colony formation in Microcystis was determined based on growth rates and extracellular polysaccharide (EPS) contents. The colony size of both strains increased significantly when the temperature rose from 15 to 25 °C. A further increase in temperature from 25 to 30 °C, however, reduced the colony size of M. ichthyoblabe significantly, and, in contrast, increased the colony size of M. wesenbergii. Higher phosphorus availability promoted the formation of larger colonies in both strains. In comparison, nitrogen had no significant effect on the colony size. Furthermore, although EPS was a significant contributor to the formation of large colonies in colonial Microcystis, growth rate was a dominant driving factor in this process. The findings of this study highlight that warmer temperatures and phosphorus enrichment might enhance surface Microcystis scums directly through increasing the colony size. This study also provides new insights into the mechanism of colony formation in Microcystis.

Succession of phytoplankton functional groups and their driving factors in a subtropical plateau lake.

The present study was carried out in a mesotrophic subtropical plateau lake, Erhai, located in southwest of China. The succession of phytoplankton functional groups and the environmental variables in the lake were investigated from January 2013 to December 2015. The lake had strong radiation levels and a low-temperature amplitude because of its high elevation and strongly mixed water. It was highly affected by the subtropical monsoon precipitation, and its pollution sources were from diffuse pollution caused by rainfall runoff. Altogether 112 genera, 16 functional groups and 4 predominant functional groups, LM (Microcystis), P (Melosira, Fragilaria, Closterium), T (Mougeotia), T (Psephonema aenigmaticum) and Y (Cryptomonas), were identified, and the predominant functional groups demonstrated strong seasonal variations. Group T (Mougeotia) dominated from the winter to early spring, with strong lake water mixing. Group P replaced group T (Mougeotia) as the dominate group of the phytoplankton community in the spring (March to May), with clear water and increased temperature. With the arrival of the monsoon rainy season in the summer, large amounts of external pollutants were brought into the lake via rainfall runoff, allowing group LM (Microcystis) to become dominant. Meanwhile, the intensive nutrient inputs after the rainstorm in the summer, combined with high temperatures and decreased radiation, led to the sustained growth of group LM in the autumn and even ultimately triggered Microcystis blooming. Group T (P. aenigmaticum) was a particular phytoplankton genus predominant in the autumn, which displayed a seasonal variation similar to that of group LM. This study underscores the usefulness of phytoplankton functional groups in studying phytoplankton succession in subtropical plateau lakes impacted by diffuse pollution, in which the succession of phytoplankton functional groups can be significantly affected by rainfall runoff, which altered variables such as nutrients, high temperatures and radiation.

The responses of the taxa composition of particle-attached bacterial community to the decomposition of Microcystis blooms.

The changes of taxa within the particle-attached bacterial assemblage during the decomposition of Microcystis blooms were investigated under darkness and anoxic condition in mesocosm experiments. During 14 days of darkness incubation, chlorophyll-a (Ch-a) concentration decreased from 2000 µg/L to 5 µg/L. Samples were collected on days 0, 2 and 14 for bacterial 16S rRNA analysis, based on rapid decreases in the Chl-a concentration of water column. The total bacterial community DNA was extracted and 16S rRNA genes were amplified by polymerase chain reaction, cloned and sequenced of selected samples. The results showed that the abundance of attached bacteria increased significantly, and the composition of the particle-attached bacterial communities varied temporally during the decomposition of Microcystis blooms. The bacterial assemblage appeared to be dominated by members of Bacteroidetes, Alphaproteobacteria and Betaproteobacteria. Shift of some genera of Alphaproteobacteria and Alphaproteobacteria was also observed. Additionally, we found that the family Sphingomonas, affiliated with Alphaproteobacteria, identified as a microcystin-degrading bacterium, dominated the particle-attached bacterial communities. The results from the present study, together with previously published data highlighted the need for more studies concerning the bacterial degradation process in order to trace the environmental fate of microcystins in field conditions.