Toxic Microcystis Research Articles

Hepatocyte apoptosis is triggered by hepatic inflammation in common carp acutely exposed to microcystin-LR or chronically exposed to Microcystis

Cyanobacterial blooms pose a serious threat to the survival of fish because of the hepatotoxicity of microcystins produced by toxic cyanobacteria such as Microcystis. Many studies have investigated the hepatotoxicity of microcystins in common carp, a freshwater fish distributed worldwide, but the hepatotoxicity mechanism has not been fully clarified. The present study aimed to investigate the mechanism underlying the hepatic inflammatory response and hepatocyte apoptosis induced by acute microcystin-LR exposure via intraperitoneal injection (71 μg/kg and 119 μg/kg) or gavage (357.08 μg/kg) and chronic exposure to toxic Microcystis blooms. The results of acute exposure revealed that microcystin-LR caused an increase in serum transaminase activity and increased the levels of inflammatory factors and inflammatory mediators, inducing a significant inflammatory response in the liver of common carp. Moreover, biochemical detection revealed that hepatocyte apoptosis occurred in the fish. Moreover, chronic toxic Microcystis exposure also caused hepatic inflammation and subsequent apoptosis mediated by the tumour necrosis factor-α (TNF-α) pathway and the mitochondrial pathway similar to acute exposure. Therefore, our study suggests that the inflammatory response induced by microcystin-LR exacerbates apoptosis, likely mediated by TNF-α. In summary, both acute microcystin-LR exposure and chronic toxic Microcystis exposure can cause inflammation in the liver of common carp, which subsequently triggers hepatocyte apoptosis mediated by the TNF-α pathway and the mitochondrial pathway. This study helps elucidate the mechanism of liver damage induced by cyanobacterial blooms in natural water.

Regulatory role of microcystin in the response of microcystin-producing cyanobacteria to elevated CO2: Insights from metabolic profiling

The regulatory role of microcystin in the response of microcystin-producing cyanobacteria to elevated CO2 remains poorly understood. To address this gap, this study compared the responses of wild-type toxic Microcystis PCC 7806 and its mcyB-knockout mutant to elevated CO2 using metabolomic profiling under nitrogen (N)-rich and N-poor conditions. Under N-poor conditions, elevated CO2 promoted carbohydrate synthesis and tricarboxylic acid cycle in both strains, without affecting their growth. Under N-rich conditions, both strains exhibited increased biomass with rising CO2 levels, attributed to enhanced carbohydrate synthesis, tricarboxylic acid cycle, glutamate-glutamine cycle, purine synthesis, and arginine synthesis. However, compared to the mutant, the proliferation of wild-type toxic Microcystis was less stimulated by elevated CO2. The difference was associated with its reduced activity in the pentose phosphate pathway, likely linked to microcystin synthesis. Besides, the difference was related to higher utilization of glutamate, arginine, and aspartate due to increased microcystin production, indicating the regulatory role of microcystin in the response of microcystin-producing cyanobacteria to elevated CO2. Importantly, elevated CO2 could enhance microcystin synthesis by promoting the production of carbon backbones (malonyl CoA), amino acids (including arginine, glutamate and aspartate) and methyl donors (S-adenosylmethionine) of the wild-type toxic Microcystis PCC 7806. Notably, sufficient nitrogen sources were required for increased amino acid and methyl donors synthesis at high CO2 concentration. The discovery revealed underlying mechanisms behind the potential for elevated CO2 levels to increase toxicity risk associated with Microcystis blooms.

Seasonal dynamics of toxigenic Microcystis in a large, shallow Lake Peipsi (Estonia) using microcystin mcyE gene abundance.

Large and temperate Lake Peipsi is the fourth largest lake in Europe, where the massive cyanobacterial blooms are composed mostly of Microcystis spp., which have been common for several decades now. The seasonal dynamics of potentially toxic Microcystis were studied using microscopy and quantitative polymerase chain reaction (qPCR) by assessing the microcystin-encoding microcystin synthetase gene E (mcyE) abundances. Water samples were analyzed over the lake areas, varying in depth, trophic level, and cyanobacterial composition during the growing period of 2021. The Microcystis mcyE genes were detected through the growing period (May-October), forming peak abundances in September with decreasing temperatures (8.9-11.1°C). Total phosphorus (TP) and nitrate (NO3-) were the most relevant environmental variables influencing the Microcystis biomass as well as mcyE abundances. Comparison with previous years (2011, 2012) indicated that the abundance and seasonal dynamics of toxigenic Microcystis can be highly variable between the years and lake areas, varying also in dominant Microcystis species. Contrary to expectations, based on mcyE abundances, the increased risk of toxin-producing Microcystis can occur in Peipsi through the growing period, independently of the water temperature and biomasses of Microcystis.

Physiological and transcriptome level responses of Microcystis aeruginosa and M. viridis to environmental concentrations of triclosan

The toxicity of triclosan (TCS) to various aquatic organisms has been demonstrated at environmental concentrations. However, the effects and mechanisms of TCS on toxic cyanobacteria remains largely unexplored. This study investigated the physiological and molecular variations in two representative toxic Microcystis species (M. aeruginosa and M. viridis) under exposure to TCS for 12 d. Our findings demonstrated that the median effective concentration (EC50) of TCS for both Microcystis species were close to the levels detected in the environment (M. aeruginosa: 9.62 μg L−1; M. viridis: 27.56 μg L−1). An increased level of reactive oxygen species (ROS) was observed in Microcystis, resulting in oxidative damage when exposed to TCS at concentrations ranging from 10 μg L−1 to 50 μg L−1. The photosynthetic activity of Microcystis had a certain degree of recovery capability at low concentrations of TCS. Compared to M. aeruginosa, the higher recovery capability of the photosynthetic system in M. viridis would be mainly attributed to the increased ability for PSII repair and phycobilisome synthesis. Additionally, the synthesis of microcystins in the two species and the release rate in M. viridis significantly increased under 10–50 μg L−1 TCS. At the molecular level, exposure to TCS at EC50 for 12 d induced the dysregulation of genes associated with photosynthesis and antioxidant system. The upregulation of genes associated with microcystin synthesis and nitrogen metabolism further increased the potential risk of microcystin release. Our results revealed the aquatic toxicity and secondary ecological risks of TCS at environmental concentrations, and provided theoretical data with practical reference value for TCS monitoring.

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.

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.

CyanoStrainChip: A Novel DNA Microarray Tool for High-Throughput Detection of Environmental Cyanobacteria at the Strain Level.

Detecting cyanobacteria in environments is an important concern due to their crucial roles in ecosystems, and they can form blooms with the potential to harm humans and nonhuman entities. However, the most widely used methods for high-throughput detection of environmental cyanobacteria, such as 16S rRNA sequencing, typically provide above-species-level resolution, thereby disregarding intraspecific variation. To address this, we developed a novel DNA microarray tool, termed the CyanoStrainChip, that enables strain-level comprehensive profiling of environmental cyanobacteria. The CyanoStrainChip was designed to target 1277 strains; nearly all major groups of cyanobacteria are included by implementing 43,666 genome-wide, strain-specific probes. It demonstrated strong specificity by in vitro mock community experiments. The high correlation (Pearson's R > 0.97) between probe fluorescence intensities and the corresponding DNA amounts (ranging from 1-100 ng) indicated excellent quantitative capability. Consistent cyanobacterial profiles of field samples were observed by both the CyanoStrainChip and next-generation sequencing methods. Furthermore, CyanoStrainChip analysis of surface water samples in Lake Chaohu uncovered a high intraspecific variation of abundance change within the genus Microcystis between different severity levels of cyanobacterial blooms, highlighting two toxic Microcystis strains that are of critical concern for Lake Chaohu harmful blooms suppression. Overall, these results suggest a potential for CyanoStrainChip as a valuable tool for cyanobacterial ecological research and harmful bloom monitoring to supplement existing techniques.

Bacteria-based Sodium Alginate Formulation to Control Toxic Microcystis Blooms

Cyanobacterial bloom formation in freshwaters is a major socio-economic and health concern across the globe. Presently used chemical and physical control strategies are inefficient in complete removal of blooms and chemical application often lead to secondary pollution in water. Hence, the current study aimed to develop a bacteria-based formulation to control toxic bloom-forming Microcystis aeruginosa in freshwaters. Two bacterial strains, Exiguobacterium acetylicum and Pseudomonas previously isolated from Sri Lankan freshwaters and characterized for M. aeruginosa cell lysis and their microcystin (MC) toxin degradation were used in the study. Initially, suitability of bacterial strains to develop into solid formulations was evaluated through biofilm formation, antibiotic sensitivity and strain compatibility. Both strains formed biofilms on the surface of microtiter plates indicating their ability to attach and colonize on solid surfaces. The multiple antibiotic resistance indices for both strains were below the threshold risk level (0.2) against the seven tested classes of antibiotics. This result indicates that there is a low risk in introducing these bacterial strains to the natural environment. Further, plate assay showed that the two strains were compatible to stay together showing no antagonistic effect on the growth of each other. Having fulfilled all three criteria tested, the two strains were immobilized into beads (~5 mm) prepared from sodium alginate at 1:1 ratio of 1×108 cells/mL bacterial inoculum. Different weights (1.0, 2.0, 3.0 g) of bacteria-immobilized beads were enclosed in sachets made with Cambrella synthetic fabric. They were introduced to M. aeruginosa grown in BG-11 medium (OD=0.2, 730 nm) and kept at 26 °C. During incubation, growth stimulation of M. aeruginosa was visually observed in 1.0 and 2.0 g beads-containing cultures, whereas, gradual discoloration of colonies was observed with 3.0 g of beads. Microscopic observations also proved complete disintegration of M. aeruginosa colonies and lysis of cells in discolored cultures. After 15 days, M. aeruginosa cell lysis was estimated as a measure of chlorophyll degradation. The highest (8.4%) cell lysis was observed in cultures containing 3.0 g of beads. This indicates that bacteria cell lysis activity depends on the load of bacteria-immobilized beads. Degradation of MC toxin was estimated by enzyme-linked immunosorbent assay. The highest (57.6%) MC degradation was observed in cultures with 3.0 g of beads after 15 days of incubation. The efficiency of bacteria release from beads was tested by placing sachets in sterilized water. At 15 days, bacterial count in water was x107 CFU/mL, with all loadings of immobilized-bacteria indicating highly efficient bacteria release from the formulation. In conclusion, this study highlights that bacteria-based sodium alginate formulations can be made as a source of inoculum to control M. aeruginosa growth and MC degradation. &#x0D; Keywords: Bacteria-based formulations, Cell lysis, Microcystis blooms, Microcystin

Controlling toxic Microcystis blooms: The power of a novel microalgal predator Poteriospumella lacustris in water safety improvement

Microcystis aeruginosa blooms have become a global environmental concern. In particular, microcystins (MC) produced by algal cells pose a great threat to aquatic ecosystem security and human health. Therefore, it is necessary to develop environmentally friendly biological measures to reduce bloom hazards. In this study, a heterotrophic flagellate, NY1, that could quickly eliminate M. aeruginosa and efficiently remove MC was isolated and identified as Poteriospumella lacustris. When the initial cell density ratio of P. lacustris to M. aeruginosa was 1:200, P. lacustris rapidly ingested M. aeruginosa, and the clearance rates were more than 90 % within 24 h. P. lacustris effectively removed 83.06 % of MC (more than 200 μg/L) in 24 h and 93.88 % in 72 h. Although temperatures lower than 15 °C and acidic conditions (pH 6) slightly and temporarily inhibited the ingestion and MC removal activities of P. lacustris within 24 h of treatment, it rapidly recovered and ingested more than 90 % of M. aeruginosa and eliminated 97 % of MC within 72 h at various temperatures (15–35 °C) and different pH values (pH 6–10). The fluorescence emission-excitation matrix spectra analysis and acute toxicity test showed that P. lacustris treatment reduced dissolved organic matter (DOM) and eliminated the toxicity of M. aerugionsa culture to three representative species at different trophic levels (Photobacterium phosphoreum, Daphnia magna, and Ctenopharyngodon idellus), indicating the promising potential of P. lacustris in improving water safety. In addition to M. aeruginosa, P. lacustris ingested and digested Platymonas subcordiformis, Chlorella vulgaris, and Synechocystis sp. This study provides a natural and biological solution to global Microcystis blooms by using the grazing effect of P. lacustris to remove toxic cyanobacteria and eliminate MC and DOM, ultimately improving water quality and safety.

Performance and characterization of snail adhesive mucus as a bioflocculant against toxic Microcystis

Toxic Microcystis blooms are widespread in aquatic bodies, posing major threats to aquatic and human life. Recently, bioflocculants have attracted considerable attention as a promising biomaterial for Microcystis management. In search of a novel organism that can produce an efficient bioflocculant for controlling harmful algae sustainably, the native gastropod Cipangopaludina chinensis was co-cultured continuously with toxic Microcystis under different initial algal cell densities. The bioflocculation effect of snail mucus on toxic Microcystis, microcystin removal, and toxin accumulation in snails was investigated. In addition, the properties of the adhesive mucus were characterized using microscopic, X-ray diffraction, infrared spectroscopy, and polysaccharide and proteome analyses. Microcystis cells were captured and flocculated by the snail mucus; removal efficiencies of up to 89.9% and 84.8% were achieved for microalgae and microcystin-leucine arginine (MC-LR), respectively, when co-cultured with C. chinensis for only one day. After nine-day exposure, less than 5.49 µg/kg DW microcystins accumulated in the snails, indicating safety for human consumption. The snail mucus contained 104.3 µg/mg protein and 72.7 µg/mg carbohydrate, which provide several functional groups beneficial for Microcystis bioflocculation. The main monosaccharide subunits of polysaccharides are galactose, galactosamine, glucosamine, fucose, glucose, and mannose. Most of them are key components of polysaccharides in many bioflocculants. Gene Ontology analysis indicated the protein enrichment in binding processes and catalytic activity, which may account for Microcystis bioflocculation via protein binding or enzymatic reactions. The findings indicate that native C. chinensis secretes adhesive mucus that can act as bioflocculant for toxic Microcystis from ambient water and can be an effective and eco-friendly tool for Microcystis suppression.

Spatio-temporal variation of toxin-producing gene abundance in Microcystis aeruginosa from Poyang Lake.

Microcystis aeruginosa (M. aeruginosa) causes massive blooms in eutrophic freshwater and releases microcystin. Poyang Lake is the largest freshwater lake in China and has kept a mid-nutrient level in recent years. However, there is little research on microcystin production in Poyang Lake. In this study, water and sediment samples from ten sampling sites in Poyang Lake were collected from May to December in 2020, and from January to April in 2021 respectively. Microcystis genes (mcyA, mcyB, 16s rDNA) were quantified by real-time fluorescence quantitative PCR analysis, and then the spatial and temporal variation of mcy genes, physicochemical factors, and bacterial population structure in the lake was analyzed. The relationship between the abundance of mcy genes and physicochemical factors in water column was also revealed. Results indicated that the microcystin-producing genes mcyA and mcyB showed significant differences in spatial and temporal levels as well, which is closely related to the physicochemical factors especially the water temperature (p < 0.05) and the nitrogen content (p < 0.05). The abundance of mcy genes in the sediment in December affected the abundance of mcy genes in the water column in the next year, while the toxic Microcystis would accumulate in the sediment. In addition to the toxic Microcystis, we also found a large number of non-toxic Microcystis in the water column and sediment, and the ratio of toxic to non-toxic species can also affect the toxicity production of M. aeruginosa. Overall, the results showed that M. aeruginosa toxin-producing genes in Poyang Lake distributed spatially and temporally which related to the physicochemical factors of Poyang Lake.

Phytoplankton functional groups as indicators of environmental changes in weir and non-weir sections of the lower Nakdong River, Republic of Korea

The Nakdong River underwent water impoundment after eight weirs were constructed as part of South Korea's Four Major River Restoration Project from 2009 to 2012. In this study, we aimed to confirm whether the assemblage of phytoplankton based on phytoplankton functional groups (PFGs), could indicate environmental changes in the weir section (WS) and non-weir section (NWS) of the lower Nakdong River after the construction of the weir. Thus, we examined the relationships between PFGs and gradients in environmental drivers, such as physicochemical, meteorological, and hydrological variables. Environmental gradients were observed between the WS and NWS in dissolved oxygen (DO), electric conductivity (EC), biochemical oxygen demand (BOD), chemical oxygen demand (COD), total nitrogen (TN), total phosphorus (TP), dissolved total nitrogen (DTN), dissolved total phosphorus (DTP), ammonia nitrogen (NH3–N), nitrate nitrogen (NO3–N), and phosphorus (PO4–P), which were relatively higher in the WS. Seventeen PFGs were identified (A, B, C, D, E, F, G, H1, J, LM, LO, MP, P, T, W1, X1, and X2).Additionally, the LM and P groups, preferring an enriched lentic system more than other groups, were found to be the dominant PFGs that led the succession of assemblages. Traditional nutrients (N, P) and organic pollutants (BOD, COD) primarily affected the autochthonous growth of the most dominant PFGs in the WS as HRT (hydraulic retention time) increased. Furthermore, the hydrological variables associated with meteorological conditions have a synergistic effect on the composition of the major PFGs and chemical and physical variables in the WS. In other words, the WS may be a new source of inoculum that primarily determines the occurrence and maintenance of phytoplankton in the immediate downstream region (NWS). In particular, group LM (mainly potentially toxic Microcystis) developing in the upper weir impoundment is transported downstream, resulting in a high inoculation effect on further growth in the NWS during the summer monsoon season.

Changes in microcystin concentration in Lake Taihu, 13 years (2007–2020) after the 2007 drinking water crisis

Since the 2007 water crisis occurred in Lake Taihu, substantial measures have been taken to restore the lake. This study evaluates the effectiveness of these restoration activities. We examined the physicochemical parameters and the distribution of microcystin and Microcystis in both the water column and sediment during the bloom period of May 2020 to October 2020. The mean value of extracellular and intracellular microcystin content was 0.12 μg L−1 and 16.26 μg L−1, respectively. The mean value of microcystin in sediment was 172.02 ng g−1 and peaked in August. The concentration in the water and sediment was significantly lower than the historical average concentration. The abundance of toxigenic Microcystis and total Microcystis in the water column ranged from 2.61 × 102 to 2.25 × 109 copies·L−1 and 8.28 × 105 to 2.76 × 109 copies·L−1, respectively. The proportion of toxic Microcystis in the sediment ranging from 31.2% to 19.12%. The highest and lowest region was Meiliang Bay and Grass-algae type zone, respectively. The copy number of the 16S rRNA gene was 1–4 orders of magnitude higher than that of mcyA gene in populations of Microcystis, indicating that non-toxic Microcystis was the dominant form in the majority of the lake. The abundance of toxic Microcystis in the water column was positively correlated with total phosphorus, PO43--P and pH, while the water temperature played distinct role to the distribution of toxic Microcystis in sediment. Our research indicated phosphorus remains a key factor influencing the toxic Microcystis and microcystins in the water column. pH played distinct roles in the distribution of microcystins in sediment and water column. The increasing water temperature is a threat. Explicit management actions and policies, which take into account nutrient concentrations, pH, and increasing temperatures, are necessary to understand and control the distribution of microcystin and Microcystis in Lake Taihu.

Cyanobacterial effects on an aquatic keystone grazer are reshaped by presence of the herbicide atrazine

Abstract As cyanobacterial blooms and herbicide pollution, which are often detected in eutrophic waters, can separately jeopardise zooplankton populations, there is an urgent and on‐going need to understand the strength and direction of their interactive effects. This is a crucial step toward realistic risk‐evaluation of agricultural pollution in eutrophic waterbodies. In this study, we evaluated how the herbicide, atrazine (ATZ), alters the effects of cyanobacterial food on the zooplankter, Daphnia magna. We found survival time of Daphnia decreased with increasing amounts of Microcystis in their diets, and the magnitude of this cyanobacterial effect was independent of ATZ. In contrast, ATZ exposure triggered faster growth and larger body size in Daphnia fed diets containing Microcystis compared to those fed the good‐food diet. Although toxic Microcystis reduced the overall reproductive output of Daphnia, the presence of ATZ, regardless of the type of food treatment, exhibited a masking effect by further significantly decreasing Daphnia's overall reproductive output, resulting in a low level of reproduction. Finally, we found an expression trade‐off at the molecular scale between growth and reproduction genes on one side, and antioxidation gene on the other, which could account in part for ATZ's influence on Microcystis toxicity to Daphnia at maturation stage. These results demonstrated that ATZ can reshape Daphnia's responses to Microcystis, predominantly with effects on growth and reproductive traits. These trait‐dependent responses were found to be closely linked to the regulation of key metabolic pathways. Collectively, our study enhances current knowledge regarding the potential interaction between fundamental trophic levels in cyanobacteria‐dominated lakes around farmlands, and is helpful to achieve more realistic environmental risk management of agricultural pollution in eutrophic waterbodies.

Microcystin aids in cold temperature acclimation: Differences between a toxic Microcystis wildtype and non-toxic mutant

For Microcystis aeruginosa PCC 7806, temperature decreases from 26 °C to 19 °C double the microcystin quota per cell during growth in continuous culture. Here we tested whether this increase in microcystin provided M. aeruginosa PCC 7806 with a fitness advantage during colder-temperature growth by comparing cell concentration, cellular physiology, reactive oxygen species damage, and the transcriptomics-inferred metabolism to a non-toxigenic mutant strain M. aeruginosa PCC 7806 ΔmcyB. Photo-physiological data combined with transcriptomic data revealed metabolic changes in the mutant strain during growth at 19 °C, which included increased electron sinks and non-photochemical quenching. Increased gene expression was observed for a glutathione-dependent peroxiredoxin during cold treatment, suggesting compensatory mechanisms to defend against reactive oxygen species are employed in the absence of microcystin in the mutant. Our observations highlight the potential selective advantages of a longer-term defensive strategy in management of oxidative stress (i.e., making microcystin) vs the shorter-term proactive strategy of producing cellular components to actively dissipate or degrade oxidative stress agents.

Is a lower-toxicity strain of Microcystis aeruginosa really less toxic?

Some well-known hazards of blooming cyanobacteria are caused by toxic metabolites such as microcystins (MCs), though many other bioactive chemicals of unknown toxicity are present in their exudates. It is also unclear whether toxicity of cyanobacterial cells depends on growth phases in the life cycle. In this study, we compared toxicity to Daphnia magna of Microcystis aeruginosa - a common cyanobacterial species - exudates (MaE) from two MC-producing strains over both exponential growth and stationary phases in acute and chronic experiments. Specifically, we assessed mitochondrial dysfunction, oxidative stress and lipid peroxidation, and filtering activity and heartbeat rate of Daphnia exposed to MaE. All MaE treatments induced common characteristics of Microcystis toxicity including disorder in the mitochondrial membrane and aberrant heart rate. MaE from cells at stationary growth phase were more toxic than those at exponential phase. Surprisingly, the MC-lower strain had higher toxicity than MC-higher one. Microcystis at different stage of blooms may differentially affect waterfleas owing to variable MaE-induced physiological dysfunction, abundance and grazing rate. Our study suggested that Microcystis strains with lower microcystin-producing ability might release other detrimental chemicals and should not be ignored in harmful bloom monitoring.

Beyond cyanotoxins: increased Legionella, antibiotic resistance genes in western Lake Erie water and disinfection-byproducts in their finished water.

Western Lake Erie is suffering from harmful cyanobacterial blooms, primarily toxic Microcystis spp., affecting the ecosystem, water safety, and the regional economy. Continued bloom occurrence has raised concerns about public health implications. However, there has been no investigation regarding the potential increase of Legionella and antibiotic resistance genes in source water, and disinfection byproducts in municipal treated drinking water caused by these bloom events. Over 2 years, source water (total n = 118) and finished water (total n = 118) samples were collected from drinking water plants situated in western Lake Erie (bloom site) and central Lake Erie (control site). Bloom-related parameters were determined, such as microcystin (MC), toxic Microcystis, total organic carbon, N, and P. Disinfection byproducts (DBPs) [total trihalomethanes (THMs) and haloacetic acids (HAAs)] were assessed in finished water. Genetic markers for Legionella, antibiotic resistance genes, and mobile genetic elements were quantified in source and finished waters. Significantly higher levels of MC-producing Microcystis were observed in the western Lake Erie site compared to the control site. Analysis of DBPs revealed significantly elevated THMs concentrations at the bloom site, while HAAs concentrations remained similar between the two sites. Legionella spp. levels were significantly higher in the bloom site, showing a significant relationship with total cyanobacteria. Abundance of ARGs (tetQ and sul1) and mobile genetic elements (MGEs) were also significantly higher at the bloom site. Although overall abundance decreased in finished water, relative abundance of ARGs and MGE among total bacteria increased after treatment, particularly at the bloom site. The findings underscore the need for ongoing efforts to mitigate bloom frequency and intensity in the lake. Moreover, optimizing water treatment processes during bloom episodes is crucial to maintain water quality. The associations observed between bloom conditions, ARGs, and Legionella, necessitate future investigations into the potential enhancement of antibiotic-resistant bacteria and Legionella spp. due to blooms, both in lake environments and drinking water distribution systems.

Chemodiversity of Cyanobacterial Toxins Driven by Future Scenarios of Climate Warming and Eutrophication.

Climate change and eutrophication are two environmental threats that can alter the structure of freshwater ecosystems and their service functions, but we know little about how ecosystem structure and function will evolve in future scenarios of climate warming. Therefore, we created different experimental climate scenarios, including present-day conditions, a 3.0 °C increase in mean temperature, and a "heatwaves" scenario (i.e., an increase in temperature variability) to assess the effects of climate change on phytoplankton communities under simultaneous stress from eutrophication and herbicides. We show that the effects of climate warming, particularly heatwaves, are associated with elevated cyanobacterial abundances and toxin production, driven by a change from mainly nontoxic to toxic Microcystis spp. The reason for higher cyanobacterial toxin concentrations is likely an increase in abundances because under the dual pressures of climate warming and eutrophication individual Microcystis toxin-producing ability decreased. Eutrophication and higher temperatures significantly increased the biomass of Microcystis, leading to an increase in the cyanobacterial toxin concentrations. In contrast, warming alone did not produce higher cyanobacterial abundances or cyanobacterial toxin concentrations likely due to the depletion of the available nutrient pool. Similarly, the herbicide glyphosate alone did not affect abundances of any phytoplankton taxa. In the case of nutrient enrichment, cyanobacterial toxin concentrations were much higher than under warming alone due to a strong boost in biomass of potential cyanobacterial toxin producers. From a broader perspective our study shows that in a future warmer climate, nutrient loading has to be reduced if toxic cyanobacterial dominance is to be controlled.

Environmental concentrations of anionic surfactants in lake surface microlayers enhance the toxicity of Microcystis blooms: Insight from photosynthesis, interspecies competition, and MC production

Anionic surfactants represented by linear alkylbenzene sulfonate (LAS) exhibit vertical heterogeneity of concentrations in aquatic environments owing to their amphiphilic structure. Field investigations showed that the concentration of anionic surfactants (mainly LAS) in the water surface microlayer (SML) of Lake Taihu reached 580 μg/L, higher than that in the lower layer. Floating Microcystis blooms overlap in space with the high concentration of anionic surfactants in SML. However, few studies have focused on the effects of anionic surfactants (e.g., LAS) on the interspecies competition between toxic and nontoxic Microcystis. In this study, coculture and monoculture experiments were conducted with both toxic and nontoxic Microcystis species to explore how the environmental concentration of LAS regulates the dominance of toxic Microcystis and toxicity from the perspective of photosynthesis, species dominance, and MC production. The results showed that LAS concentrations above 0.267 or 0.431 mg/L (depending on light conditions) selectively promoted the photosynthetic competitive advantage of toxic Microcystis, leading to its higher population proportion in the community. Additionally, LAS concentrations above 0.5 mg/L induced the synthesis and release of microcystins (MCs). The results of chlorophyll fluorescence analysis, electron microscopy and transcriptome sequencing suggested that compared with nontoxic Microcystis, toxic Microcystis can better resist LAS stress by dissipating excess light, maintaining an intact membrane structure and maintaining cellular homeostasis. Transcriptome sequencing revealed that the photosynthetic damage of nontoxic Microcystis might be attributed to the impacts of LAS on the absorption and assimilation of nitrogen, which finally resulted in the degradation of phycobilisomes. This study can provide novel insight for establishing standards and safety management of wastewater discharge.

Temperature-Dependent Growth Characteristics and Competition of Pseudanabaena and Microcystis

Global warming has been considered to accelerate the expansion of cyanobacterial blooms, which are frequently composed of the bloom-forming genera, Microcystis and Pseudanabaena, in freshwater ecosystems worldwide. Nonetheless, the impact of changes due to toxin production or lack thereof on the growth of co-existing strains, both arising from increasing temperature, has remained unknown to date. We conducted competition experiments involving toxic Microcystis PCC7806, a non-toxic mcyB− mutant, and two strains of Pseudanabaena (TH-1 and DC-1) identified as P. mucicola. In mono-culture, the specific growth ratio of Pseudanabaena increased; nevertheless, the maximum cell density declined with increasing temperature. The maximum growth ratios of Pseudanabaena TH-1 and Pseudanabaena DC-1 were 0.512 day−1 in the 30 °C group and 0.440 day−1 in the 35 °C group, respectively. The optimum temperature for the growth of Pseudanabaena was 25 °C. Remarkably, there was no significant disparity in the maximum cell density of Microcystis PCC7806 and the mcyB− mutant across varied temperature groups, even though their maximum growth rates differed marginally, reaching 0.280 day−1 and 0.306 day−1 in the 30 °C group, respectively. In co-cultures, the growth of Pseudanabaena TH-1 was uniformly inhibited, whereas that of DC-1 was somewhat influenced by co-culturing with toxic and non-toxic Microcystis, except for the 35 °C group, where inhibition was absent amongst DC-1 and Microcystis. Moreover, the growth of Microcystis was promoted with a co-culture of TH-1 in the 20 °C groups. Conversely, the co-culture of Microcystis with Pseudanabaena DC-1 led to the inhibition of the former in the 30 °C and 35 °C groups. With a Lotka–Volterra competition model, the results showed that Microcystis dominated or co-existed with Pseudanabaena, conforming to expectations from the wild.