Concentrations Of microcystin-LR Research Articles

Feasibility and mechanism of removing Microcystis aeruginosa and degrading microcystin-LR by dielectric barrier discharge plasma

Harmful cyanobacterial bloom is one of the serious environmental problems worldwide. Microcystis aeruginosa is a representative harmful alga in cyanobacteria bloom. It is of great significance to develop new technologies for the removal of Microcystis aeruginosa and microcystins. The feasibility and mechanism of removing microcystis aeruginosa and degrading microcystins by dielectric barrier discharge (DBD) plasma were studied. The suitable DBD parameters obtained in this study are DBD (41.5 W, 40 min) and DBD (41.5 W, 50 min), resulting in algae removal efficiency of 77.4% and 80.4%, respectively; scanning electron microscope and LIVE-DEATH analysis demonstrate that DBD treatment can disrupt cell structure and lead to cell death; analysis of elemental composition and chemical state indicated that there are traces of oxidation of organic nitrogen and organic carbon in microcystis aeruginosa; further intracellular ROS concentration and antioxidant enzyme activity analysis confirm that DBD damage microcystis aeruginosa through oxidation. Meanwhile, DBD can effectively degrade the microcystin-LR released after cell lysis, the extracellular microcystin-LR concentration in the DBD (41.5 W) group decreased by 88.7% at 60 min compared to the highest concentration at 20 min; further toxicity analysis of degradation intermediates indicated that DBD can reduce the toxicity of microcystin-LR. The contribution of active substances to the inactivation of microcystis aeruginosa is eaq− >•OH > H2O2 > O3 > 1O2 >•O2− >ONOO−, while on the degradation of microcystin-LR is eaq− >•OH > H2O2 > O3 >•O2− >1O2 > ONOO−. The application of DBD plasma technology in microcystis aeruginosa algae removal and detoxification has certain prospects for promotion and application.

Reusable and Practical Biocomposite Based on Sphingopyxis sp. YF1 and Polyacrylonitrile-Based Carbon Fiber for the Efficient Bioremediation of Microcystin-LR-Contaminated Water.

Microbial degradation is a cost-effective and environmentally friendly method for removing microcystin-LR (MC-LR). However, the application of free bacteria has limitations due to low operational stability and difficulties in recovery. In a previous study, our group successfully isolated a highly efficient MC-LR-degrading bacterium, Sphingopyxis sp. YF1, from Taihu. To enhance its practical potential in addressing MC-LR-contaminated water pollution, a novel biological material named polyacrylonitrile-based carbon fiber @Sphingopyxis sp. YF1 (PAN-CF@YF1) was synthesized. The immobilization conditions of strain Sphingopyxis sp. YF1 on PAN-CF surfaces were optimized using Box-Behnken design and response surface methodology (RSM), which turned out to be an optimal pH of 7.6 for the culture medium, a ratio of 0.038 g of supporting materials per 100 mL of culture media, and an incubation time of 53.4 h. The resultant PAN-CF@YF1 showed a great degradation effect both for low and high concentrations of MC-LR and exhibited satisfactory cyclic stability (85.75% after six cycles). Moreover, the application of PAN-CF@YF1 in the bioreactors demonstrated effective and sustainable MC-LR removal, with a removal efficiency of 78.83% after three consecutive treatments. Therefore, PAN-CF@YF1 with high degradation activity, environmental compatibility, straightforward preparation, and recyclability shows significant application potential for the bioremediation of MC-LR-contaminated water bodies.

Development of an Integrated Multi-Method Electrochemical Biosensor for Rapid-on-Site Detection and Quantification of Cyanotoxins

The accumulation of Cyanotoxins (CTXs) in water reservoirs is an emerging global concern. Exposure to microcystin-LR (MC-LR), among the most common CTXs, is associated with hepatotoxicity and carcinogenesis and leads to both acute and chronic damages. Stringent regulation has recently been imposed by the World Health Organization, limiting the allowed MC-LR concentration in drinking water to <1 μg/L. Current methods for MC-LR detection require expensive equipment, trained personnel, and labor-intensive preparation. Therefore, there is an urgent need for an affordable, on-site diagnostic tool providing a rapid quantitative determination of MC-LR in various water types. In this study, we have developed an electrochemical (EC) biosensor that comprised of a miniaturized and multiplexed microchips, functionalized with anti-MC-LR antibodies, and MC-LR detection is based on the transduction of bimolecular binding into an electrochemical signal. The transduction method applied is based on the comparison of two (2) electrochemical techniques: a label-free Electrochemical Impedance Spectroscopy (EIS) a labeled Electrochemical Immunoassay (ECI) detection of MC-LR in <10 min. The former, the EIS-based detection method, demonstrated the feasibility of detecting ultra-low (3 ppt or ng/L) MC-LR concentrations, and preliminary results also showed that the biochip demonstrated its use in rapid quantitative detection of microcystins in raw cyanobacterial cultures. However, the latter, ECI-based detection method showed a detection limit of 3µg/L (3ppb) of purified MC-LR concentrations, which is an order of magnitude less sensitive than the EIS-based detection method. These LODs obtained from the two methods are superior to currently employed solid-phase immunoassays and other analytical methods. Based on our findings, we anticipate that in the near future electrochemical biosensors would be an essential tool in water monitoring and environmental diagnostics. Figure 1

Hepatotoxicity of cyanotoxin microcystin-LR in human: Insights into mechanisms of action in the 3D culture model Hepoid-HepaRG

Microcystin-LR (MC-LR) is a potent hepatotoxin produced by harmful cyanobacterial blooms (CyanoHABs). MC-LR targets highly differentiated hepatocytes expressing organic anion transporting polypeptides OATP1B1 and OATP1B3 that are responsible for hepatocellular uptake of the toxin. The present study utilized an advanced 3D in vitro human liver model Hepoid-HepaRG based on the cultivation of collagen-matrix embedded multicellular spheroids composed of highly differentiated and polarized hepatocyte-like cells. 14-d-old Hepoid-HepaRG cultures showed increased expression of OATP1B1/1B3 and sensitivity to MC-LR cytotoxicity at concentrations >10 nM (48 h exposure, EC20 = 26 nM). MC-LR induced neither caspase 3/7 activity nor expression of the endoplasmic reticulum stress marker gene BiP/GRP78, but increased release of pro-inflammatory cytokine IL-8, indicating a necrotic type of cell death. Subcytotoxic (10 nM) and cytotoxic (≥100 nM) MC-LR concentrations disrupted hepatocyte functions, such as xenobiotic metabolism phase-I enzyme activities (cytochrome P450 1A/1B) and albumin secretion, along with reduced expression of CYP1A2 and ALB genes. MC-LR also decreased expression of HNF4A gene, a critical regulator of hepatocyte differentiation and function. Genes encoding hepatobiliary membrane transporters (OATP1B1, BSEP, NTCP), hepatocyte gap junctional gene connexin 32 and the epithelial cell marker E-cadherin were also downregulated. Simultaneous upregulation of connexin 43 gene, primarily expressed by liver progenitor and non-parenchymal cells, indicated a disruption of tissue homeostasis. This was associated with a shift in the expression ratio of E-cadherin to N-cadherin towards the mesenchymal cell marker, a process linked to epithelial-mesenchymal transition (EMT) and hepatocarcinogenesis. The effects observed in the human liver cell in vitro model revealed mechanisms that can potentially contribute to the MC-LR-induced promotion and progression of hepatocellular carcinoma (HCC). Hepoid-HepaRG cultures provide a robust, accessible and versatile in vitro model, capable of sensitively detecting hepatotoxic effects at toxicologically relevant concentrations, allowing for assessing hepatotoxicity mechanisms, human health hazards and impacts of environmental hepatotoxins, such as MC-LR.

Responses of &lt;i&gt;Unio pictorum&lt;/i&gt; to the Presence of Toxic and Non-Toxic Strains of &lt;i&gt;Microcystis aeruginosa&lt;/i&gt;

In order to assess the impact of cyanobacteria on mollusks under experimental conditions, the interaction of toxic and non-toxic strains of cyanobacteria Microcystis aeruginosa (Kützing) Kützing and bivalve mollusks Unio pictorum (L., 1758) was studied. Cyanobacteria have a negative effect on bivalve mollusks: 40% death of mollusks and deterioration of their adaptive capacity were recorded when co-cultivated with M. aeruginosa at a high cell concentration. At the same time, there was no difference in the mortality of mollusks incubated with toxic and non-toxic cyanobacteria. A decrease in the content of microcystin LR in the presence of bivalves was revealed. No statistically significant increase in the number of cyanobacteria in the water was noted after transit passage through the digestive system of bivalves.

Nature-Based Solution to Eliminate Cyanotoxins in Water Using Biologically Enhanced Biochar.

Climate change and high eutrophication levels of freshwater sources are increasing the occurrence and intensity of toxic cyanobacterial blooms in drinking water supplies. Conventional water treatment struggles to eliminate cyanobacteria/cyanotoxins, and expensive tertiary treatments are needed. To address this, we have designed a sustainable, nature-based solution using biochar derived from waste coconut shells. This biochar provides a low-cost porous support for immobilizing microbial communities, forming biologically enhanced biochar (BEB). Highly toxic microcystin-LR (MC-LR) was used to influence microbial colonization of the biochar by the natural lake-water microbiome. Over 11 months, BEBs were exposed to microcystins, cyanobacterial extracts, and live cyanobacterial cells, always resulting in rapid elimination of toxins and even a 1.6-1.9 log reduction in cyanobacterial cell numbers. After 48 h of incubation with our BEBs, the MC-LR concentrations dropped below the detection limit of 0.1 ng/mL. The accelerated degradation of cyanotoxins was attributed to enhanced species diversity and microcystin-degrading microbes colonizing the biochar. To ensure scalability, we evaluated BEBs produced through batch-scale and continuous-scale pyrolysis, while also guaranteeing safety by maintaining toxic impurities in biochar within acceptable limits and monitoring degradation byproducts. This study serves as a proof-of-concept for a sustainable, scalable, and safe nature-based solution for combating toxic algal blooms.

Occurrence of Harmful Algal Blooms in Freshwater Sources of Mindu and Nyumba ya Mungu Dams, Tanzania.

Harmful algal blooms (HABs) pose a significant threat to aquatic ecosystems and human health due to the production of toxins. The identification and quantification of these toxins are crucial for water quality management decisions. This study used DNA analysis (PCR techniques) to identify toxin-producing strains and liquid-chromatography-tandem mass spectrometry (LC-MS/MS) to quantify microcystins in samples from Mindu and Nyumba ya Mungu Dams in Tanzania. The results showed that HABs were detected in both dams. The BLAST results revealed that the 16S gene sequences of uncultured samples were very similar to an Antarctic cyanobacterium, Leptolyngbya sp, Anabaena sp, and Microcystis aeruginosa. Sequences of the cultured samples were most similar to Nodularia spumigena, Amazoninema brasiliense, Anabaena sp, and Microcystis aeruginosa. Further analyses showed that the nucleotide sequence similarity of uncultured isolates from this study and those from the GenBank ranged from 85 to 100%. For cultured isolates from this study and others from the GenBank, nucleotide identity ranged from 81 to 100%. The molecular identification of Microcystis aeruginosa confirmed the presence of HABs in both Mindu and Nyumba ya Mungu Dams in Tanzania. At Mindu Dam, the mean concentrations (± standard deviation) of microcystin-LR, -RR, and -YR were 1.08 ± 0.749 ppm, 0.120 ± 0.0211 ppm, and 1.37 ± 0.862 ppm, respectively. Similarly, at Nyumba ya Mungu Dam, the concentrations of microcystin-LR, -RR, and -YR were 1.07 ± 0.499 ppm, 0.124 ± 0.0224 ppm, and 0.961 ± 0.408 ppm, respectively. This paper represents the first application of PCR and LC-MS/MS to study microcystins in small freshwater reservoirs in Tanzania. This study confirms the presence of toxin-producing strains of Microcystis aeruginosa in both dams and also provides evidence of the occurrence of microcystins from these strains. These findings contribute in improving the monitoring of HABs contamination and their potential impact on water quality in Tanzanian reservoirs.

Management of the Risk of Blue-Green Algae Blooms in the Iskar Dam for Drinking Water Quality of Sofia City

The introduction of systems for ensuring quality and safe drinking water and risk management is the World Health Organization (WHO) good practice introduced as a legislative requirement for all water supply organizations (Directive EU 2020/2184) on the quality of water intended for human consumption. A specific object of this research is the implemented risk assessment and management system, part of the drinking water safety plan for Sofia city. The water supply system from the water source to the end user, operated by Sofiyska voda JSC and part of Veolia, was analyzed. The hazards and risk events were reviewed for each component of the system—the catchment, water source, supply water pipes, drinking water treatment plants (DWTPs) and distribution water network. The results of the risk evaluation show that the risk of an algal bloom falls from medium risk, with a value of 36 in 2018. This risk was new for the Iskar Dam—the main water source for Sofia city. The purpose of the current study is to analyze the risk of blue-green algae blooming and the potential release of toxins in the Iskar Dam. A data analysis of phytoplankton counts, temperature of the water in the dam, precipitation, algal diversity and quantity of microcystins for a period of three years shows that there is a low probability of algae blooming. The concentration of microcystin LR was below 0.6 µg/L and of microcystin YR was below 0.1 µg/L. The highest phytoplankton count was 70,000 cells/mL in 2018, and it decreased to 30,000 cells/mL in 2020. The impact of the risk of “blue-green algae blooms in the Iskar Dam” is assessed as moderate (3) and that has not changed between 2018 and 2022. The likelihood of occurrence of this risk decreased from possible (3) to unlikely (2) after risk reassessment, and the current control effectiveness changed from weakly controlled (4) in 2018 to well controlled (2) in 2022, because more risk management actions were included in Sofiyska voda JSC. The results of the risk evaluation show that the risk of an algal bloom falls into the low-risk category with a value of 12, after measures were considered and a reassessment. As a preventive measure for better management and risk minimization, it is necessary to continue monitoring the phytoplankton count, species composition, concentration of algae toxins (microcystin LR) and nutrients in raw water from the Iskar Dam. This research has added value to the global database for surface water sources intended for human consumption and is focuses on a potential problem, valid for agricultural as well as for urban territories rich in diffused sources of pollution. The specific risk evaluation and management steps could be used and adapted by other water supply companies.

Phytoplankton interspecific interactions modified by symbiotic fungi and bacterial metabolites under environmentally relevant hydrogen peroxide concentrations stress

Hydrogen peroxide (H2O2), which accumulates in water and triggers oxidative stress for aquatic microbes, has been shown to have profound impacts on planktonic microbial community dynamics including cyanobacterial bloom formation. Yet, potential effects of H2O2 on interspecific relationships of phytoplankton-microbe symbiotic interactions remain unclear. Here, we investigated effects of environmentally relevant H2O2 concentrations on interspecific microbial relationships in algae-microbe symbiosis. Microbes play a crucial role in the competition between M. aeruginosa and Chlorella vulgaris at low H2O2 concentrations (∼400 nM), in which fungi and bacteria protect Microcystis aeruginosa from oxidative stress. Moreover, H2O2 stimulated the synthesis and release of extracellular microcystin-LR from Microcystis aeruginosa, while intracellular microcystin-LR concentrations remained at a relatively constant level. In the presence of H2O2, loss of organoheterocyclic compounds, organic acids and ketones contributed to the growth of M. aeruginosa, but the reduction of vitamins inhibited it. Regulation of interspecific relationships by H2O2 is achieved by its action on fungal species and bacterial secretory metabolites. This study explored the response of phytoplankton interspecific relationships in symbiotic phytoplankton-microbe interactions to environmentally relevant H2O2 concentrations stress, providing a theoretical basis for understanding the formation of harmful-algae blooming and impact of photochemical properties of water on aquatic ecological safety and stability.

Immunotoxicity of cynobacterial toxin Microcystin-LR is mitigated by Quercitin and himalaya tonic Liv52

Microcystin-LR (MC-LR) has received worldwide concern for its hepatotoxicity with maximum acceptable daily intake of 0.0015 mg/L (1.5 μg/L) [Federal-Provinicial-Territorial-Committee-on-drinking-water-2002]. Comprehensive immunotoxicity data is still deficient with MC-LR. To curb the menace of MC-LR, Quercitin (QE), himalaya made hepatotonic Liv52 were studied. To investigate the immunotoxic properties of MC-LR, QE and Liv52, primary splenocyte cells prepared, cultured, and immunoproliferation assay with mitogens lipopolysaccharide (LPS) or concanavalin A, (Con A) was done for, immunophenotyping, cell cycle and apoptotic studies. In current study, we have divided the splenocytes into 4 groups, i.e., Group I: Normal saline, Group II: MC-LR (0.1 μM), Group III: MC-LR (0.1 μM) + QE (20 μM), and Group IV: MC-LR (0.1 μM) + Liv52 (25 μg/ml) and treated with maximum < CC50 concentration. MC-LR enhanced proliferation of Con A and LPS stirred splenocytes at 24 h, whereas QE and Liv52 both act as antimitogenic. With combined mixture of MC-LR + QE, a significant increase in proliferation compared to mitogen or MC-LR was observed. MC-LR down-regulated expression of CD19+, CD3e+, CD4+, CD8+, (1.05%), (18.9%), (8.9%), and (7.8%) respectively in comparison to Group I. Down-regulation of 10% and 28% is observed in CD19+ and CD4+ populations with MC-LR and QE. The Liv52 addition concealed MC-LR adverse properties in most effective way. MC-LR induced G1-phase significant declined cell cycle arrest at S phase (9.26%) and G2/M phase (26.31%) was observed. QE and Liv52 mask the activity of MC-LR. Further apoptotic study revealed that MC-LR treatment decreases late apoptotic cells compared to control with no significant change in live and early apoptotic cells. Although QE increased live cells and Liv52 significantly increased late apoptotic cells, these results suggest that a <CC50 concentration (0.1 μM) of MC-LR may have immunosuppressive activity. Liv52 is found to be a more potent drug to counteract this cyanotoxin.

Development of a chitosan-multi-walled carbon nanotubes composite for application in solid-phase adsorption toxin tracking of microcystins

Contamination of water and food with cyanotoxins poses human health risks, and hence the need for sensitive early warning tools to monitor these in water. A composite of glutaraldehyde-crosslinked chitosan and multi-walled carbon nanotubes (ChMWCNTs) was synthesised and tested for potential use as a solid-phase adsorption toxin tracking (SPATT) adsorbent for monitoring microcystins (MCs) in fresh water. The composite was characterised by Fourier transform infrared spectroscopy, Brunauer–Emmett–Teller theory and scanning electron microscopy. Batch adsorption experiments to assess the effect of contact time, adsorbent dosage and initial microcystin-LR (MC-LR) concentration were conducted. The composite was found to be efficient in adsorbing MC-LR, showing 97% removal and a maximum adsorption capacity of 4.639 μg/g under optimised conditions of 5 μg/L of MC-LR, adsorbent dose of 0.03 g/5 mL and 30 min contact time. The adsorption kinetics were better explained by a pseudo-second-order model, inferring chemisorption adsorption. The isotherm data better fitted the Langmuir isotherm model, thus inferring monolayer surface adsorption. For desorption, 100% methanol was the most effective, with an efficiency of 84.71%. The composite effectively adsorbed and desorbed three congeners of MCs (–LR, –RR and –YR) when tested in raw dam water, regardless of its lower maximum adsorption capacity compared to those of other adsorbents used for similar purposes. Significance: Monitoring of microcystins is problematic in large reservoirs and rivers. Chitosan can be crosslinked and modified to enhance its adsorption properties. Composites of chitosan and carbon nanotubes efficiently adsorb and desorb microcystins. This study is possibly the first to apply a chitosan-based sorbent in solid-phase toxin tracking (SPATT) to be used as an early warning tool in passive monitoring of microcystins in water resources. Open data set: https://doi.org/10.6084/m9.figshare.20992291.v1

Combined toxic effects of perfluorooctanoic acid and microcystin-LR on submerged macrophytes and biofilms

Perfluorooctanoic acid (PFOA) and microcystin-LR (MCLR) are pervasive pollutants in surface waters that induce significant toxic effects on aquatic organisms. However, the combined environmental risk of PFOA and MCLR remains unclear. To assess the toxic effects of PFOA and MCLR on submerged macrophytes and biofilms, Vallisneria natans was exposed to different concentrations of PFOA and MCLR (0.01, 0.1, 1.0 and 10.0 μg L−1). Vallisneria natans was sensitive to high concentrations of MCLR (10 μg L−1): plants exposed to 10 μg L−1 of MCLR measured a biomass of 3.46 g, which was significantly lower than the 8.71 g of the control group. Additionally, antagonistic interactive effects were observed in plants exposed to combined PFOA and MCLR. Exposure to these pollutants adversely affected photosynthesis of the plants and triggered peroxidation that promoted peroxidase, superoxide dismutase and catalase activities, and increased malondialdehyde and glutathione concentrations. The total chlorophyll content was lower in the highest concentration of the combined treatment group (0.443 mg g−1) than in the control group (0.534 mg g−1). Peroxidase activity increased from 662.63 U mg−1 Pr to 1193.45 U mg−1 Pr with increasing PFOA concentrations. Metabolomics indicated that the stress tolerance of Vallisneria natans was improved via altered fatty acid metabolism, hormone metabolism and carbon metabolism. Furthermore, PFOA and MCLR influenced the abundance and structure of the microbial community in the biofilms of Vallisneria natans. The increased contents of autoinducer peptide and N-acylated homoserine lactone signaling molecules indicated that these pollutants altered the formation and function of the biofilm. These results expand our understanding of the combined effects of PFOA and MCLR in aquatic ecosystems.

Sorption and Biodestruction of Microcystin-LR by &lt;i&gt;Penicillium verrucosum&lt;/i&gt; CP4 Strain Isolated from the Bottom Sediments of Sestroretsky Razliv Lake

The strain of fungus СР4 capable of degrading microcystin – LR (MC-LR) was isolated from the bottom sediments of Sestroretsky Razliv Lake. Based on DNA ITS sequencing and morphological analysis, the CP4 strain was identified as Penicillium verrucosum. The decrease in the content of MC-LR during the cultivation of strain СР4 from 0.64 μg/mL to 0.31 μg/mL occurs mainly due to biodegradation and, to a lesser extent, due to the sorption of the toxin by fungal cells. The method of biotesting (Daphnia magna) showed a decrease in the toxicity of the culture liquid in the process of MC-LR biodegradation by the strain СР4. The obtained results allow us to consider Penicillium verrucosum СР4 as a promising strain for mycoremediation of water bodies contaminated with microcystins.

Cyanotoxins Increase Cytotoxicity and Promote Nonalcoholic Fatty Liver Disease Progression by Enhancing Cell Steatosis.

Freshwater prokaryotic cyanobacteria within harmful algal blooms produce cyanotoxins which are considered major pollutants in the aquatic system. Direct exposure to cyanotoxins through inhalation, skin contact, or ingestion of contaminated drinking water can target the liver and may cause hepatotoxicity. In the current study, we investigated the effect of low concentrations of cyanotoxins on cytotoxicity, inflammation, modulation of unfolded protein response (UPR), steatosis, and fibrosis signaling in human hepatocytes and liver cell models. Exposure to low concentrations of microcystin-LR (MC-LR), microcystin-RR (MC-RR), nodularin (NOD), and cylindrospermopsin (CYN) in human bipotent progenitor cell line HepaRG and hepatocellular carcinoma (HCC) cell lines HepG2 and SK-Hep1 resulted in increased cell toxicity. MC-LR, NOD, and CYN differentially regulated inflammatory signaling, activated UPR signaling and lipogenic gene expression, and induced cellular steatosis and fibrotic signaling in HCC cells. MC-LR, NOD, and CYN also regulated AKT/mTOR signaling and inhibited autophagy. Chronic exposure to MC-LR, NOD, and CYN upregulated the expression of lipogenic and fibrosis biomarkers. Moreover, RNA sequencing (RNA seq) data suggested that exposure of human hepatocytes, HepaRG, and HCC HepG2 cells to MC-LR and CYN modulated expression levels of several genes that regulate non-alcoholic fatty liver disease (NAFLD). Our data suggest that low concentrations of cyanotoxins can cause hepatotoxicity and cell steatosis and promote NAFLD progression.

Enhancing Biocontrol of Harmful Algae Blooms: Seasonal Variation in Allelopathic Capacity of Myriophyllum aquaticum

Myriophyllum aquaticum has shown potential allelopathic effects for the biocontrol of cyanobacteria and cyanotoxins. However, the composition of allelochemicals and their biological effects may be influenced by seasonal changes. In this study, we investigated the impact of aqueous extracts of M. aquaticum collected in different seasons on the growth of Microcystis aeruginosa and the concentration of microcystin-LR. Plant samples were extracted using ultrasound cycles in aqueous solutions, and extracts at varying concentrations (0.1, 10, and 100 mg/L) and a control treatment were inoculated with M. aeruginosa, and cell growth was analyzed using a Neubauer chamber. Photosynthetic pigment quantification was used to measure physiological effects and liquid chromatography was used to evaluate the microcystin-LR concentrations. The extracts of plants collected during autumn and winter exhibited higher inhibition of M. aeruginosa growth and a reduction in photosynthetic pigments compared to those collected during spring and summer. These results can be explained by the higher presence of phenolic compounds in the composition of extracts from autumn and winter. Microcystin-LR concentrations were decreased at 10 and 100 mg/L, with the highest efficiency observed in autumn, while spring showed lower efficiency. Our findings suggest that M. aquaticum extracts have inhibitory potential on M. aeruginosa, particularly during the autumn season, making them a promising nature-based solution for the biocontrol of harmful algal blooms.

Graphene as a rational interface for enhanced adsorption of microcystin-LR from water

Cyanotoxins such as microcystin-LR (MC-LR) represent a global environmental threat to ecosystems and drinking water supplies. The study investigated the direct use of graphene as a rational interface for removal of MC-LR via interactions with the aromatic ring of the ADDA1 chain of MC-LR and the sp2 hybridized carbon network of graphene. Intra-particle diffusion model fit indicated the high mesoporosity of graphene provided significant enhancements to both adsorption capacities and kinetics when benchmarked against microporous granular activated carbon (GAC). Graphene showed superior MC-LR adsorption capacity of 75.4 mg/g (Freundlich model) compared to 0.982 mg/g (Langmuir model) for GAC. Sorption kinetic studies showed graphene adsorbs 99% of MC-LR in 30 min, compared to zero removal for GAC after 24 hr using the same MC-LR concentration. Density functional theory (DFT), calculations showed that postulated π-based interactions align well with the NMR-based experimental work used to probe primary interactions between graphene and MC-LR adduct. This study proved that π-interactions between the aromatic ring on MC-LR and graphene sp2 orbitals are a dominant interaction. With rapid kinetics and adsorption capacities much higher than GAC, it is anticipated that graphene will offer a novel molecular approach for removal of toxins and emerging contaminants with aromatic systems.

Sulfur Quantum Dot-Based Electrochemiluminescence Resonance Energy Transfer System for a Dual-Mode Ultrasensitive Detection of MC-LR Using Ag+ as a Signal Amplification Switch.

Improving the sensitivity and accuracy of bioimmunoassays has been the focus of research into the development of electrochemiluminescence (ECL) sensing platforms, as this is a critical factor in their application to practical analysis. In this work, an electrochemiluminescence-electrochemistry (ECL-EC) dual-mode biosensing platform based on an "off-on-super on" signals pattern strategy was developed for the ultrasensitive detection of Microcystin-LR (MC-LR). In this system, sulfur quantum dots (SQDs) are a novel class of ECL cathode emitter with almost no potentially toxic effects. The sensing substrate is made from rGO/Ti3C2Tx composites, whose huge specific surface area greatly reduces the possibility of aggregation-caused quenching of SQDs. The ECL detection system was constructed based on the ECL-resonance energy transfer (ERET) strategy, where methylene blue (MB) with an ECL receptor function was bound to the aptamer of MC-LR by electrostatic adsorption and the center actual distance between the donor and the acceptor was calculated to be 3.84 nm, which was verified to be in accordance with the ERET theory. Meanwhile, the introduction of Ag+ as an ECL signal-amplifying molecule greatly improved the sensitivity of sensing analysis. Based on the specific binding of MC-LR to the aptamer, the concentration of MC-LR was found to have a positive correlation with the ECL signal. Also, EC detection was realized with the benefit of the excellent electrochemical properties of MB. The dual-mode biosensor greatly improves the confidence of the detection, examination areas of 0.001-100 pg/mL with MC-LR for ECL and EC were obtained, and the detection limits are 0.17 and 0.24 pg/mL, respectively.

Flocculation and lysis of Microcystis aeruginosa by Paebubacillus sp. A9 and inhibition of microcystin release

Algae such as Microcystis aeruginosa, which are involved in cyanobacterial harmful algal blooms (CyanoHABs), pose a serious threat to aquatic organisms and human health, and have become a severe environmental problem. In the search for efficient, eco-friendly and safe approaches to control CyanoHABs, we evaluated the algicidal potential of Paebubacillus sp. A9 (A9) cells, supernatants and cultures with flocculation properties against M. aeruginosa. The results showed that different fractions of A9 exhibited algicidal activity in a dose- and time-dependent manner. Initially, polysaccharides with a series of carboxyl groups released by A9 caused flocculation of M. aeruginosa cells, which was followed by the lysis of algal cells by the algicidal active compounds present in the A9 culture. The algicidal process activated the antioxidant system of M. aeruginosa, as evidenced by a dramatic increase in the activities of catalase (CAT) and superoxide dismutase (SOD) enzyme. Simultaneously, photosynthetic pigments (Chl a), photosynthetic efficiency (Fv/Fm) and maximum relative photosynthetic electron transfer rate (ETRmax) were significantly decreased, indicating the impairment of the photosynthetic system of M. aeruginosa by A9 culture. Notably, inoculation with 6% A9 culture reduced the microcystin-LR (MC-LR) concentration by 90.65% compared to the control. Further analysis of fluorescent dissolved organic contaminants, disinfection by-product precursors, and nitrogen and phosphorus nutrients in the algal culture, confirmed the environmentally safety of the A9-mediated algicidal process. Therefore, Paebubacillus sp. A9 be a potential candidate for controlling M. aeruginosa-induced CyanoHABs through its ”flocculation–lysis–degradation” mechanism, coupled with nutrient regulation.

Acute toxic effects of microcystin-LR on crayfish (Procambarus clarkii): Insights from antioxidant system, histopathology and intestinal flora.

To study the toxic effects of microcystin-LR (MC-LR) on crayfish, adult male Procambarus clarkii were exposed to different concentrations of MC-LR for 96h. In the meantime, the accumulation characteristics of MC-LR and the alternations of antioxidant system, histopathology and intestinal flora of P. clarkii were investigated. The results demonstrated that the hepatopancreas, gills and intestines of P. clarkii could effectively accumulate MC-LR. Antioxidant-related genes such as Mn-sod, cat, gst, gpx, mt and hsp70 showed different expression trends in different organs to respond to MC-LR-induced oxidative stress. MC-LR led to histological changes in the hepatopancreas, gills and intestines, thus affecting their corresponding physiological functions. Additionally, the abundances of bacterial phyla including Firmicutes and Planctomycetes and genera including Dysgonomonas, Brevundimonas and Anaerorhabdus in the intestine were significantly changed after MC-LR exposure, and the disruption of intestinal flora might further cause abnormal intestinal microbial metabolism and genetics in P. clarkii. This study provides novel mechanistic insights into the toxic impacts of microcystins on aquatic crustaceans. HIGHLIGHTS: • MC-LR was significantly accumulated in the hepatopancreas, gills and intestines of P. clarkii. • MC-LR induced the differential expression of antioxidant-related genes of P. clarkii. • MC-LR caused histological alterations in the hepatopancreas, gills and intestines of P. clarkii. • MC-LR affected the intestinal microbial composition and function of P. clarkii.

Effect of butachlor on Microcystis aeruginosa: Cellular and molecular mechanisms of toxicity

The rapid development of agriculture increases the release of butachlor into aquatic environments. As a dominant species causing cyanobacterial blooms, Microcystis aeruginosa (M. aeruginosa) can produce microcystin and poses threats to aquatic ecosystems and human health. However, the impact of butachlor on M. aeruginosa remains unclarified. Therefore, the physiochemical responses of M. aeruginosa to butachlor were investigated, and the relevant underlying molecular mechanism was highlighted. There were no significant changes (P > 0.05) in the growth and physiology of M. aeruginosa at the low concentrations of butachlor (0–0.1 mg/L), which evidenced a high level of butachlor tolerance in Microcystis aeruginosa. For the high concentrations of butachlor (4–30 mg/L), the inhibition of photosynthetic activity, disruption of cell ultrastructure, and oxidative stress were dominant toxic effects on M. aeruginosa. Additionally, the impaired cellular integrity and lipid peroxidation may be attributed to the substantial elevations of extracellular microcystin-LR concentration. Downregulation of genes associated with photosynthesis, energy metabolism, and oxidative stress was inferred to be responsible for the growth suppression of M. aeruginosa in 30 mg/L butachlor treatment. The upregulation of gene sets involved in nitrogen metabolism may illustrate the specific effort to sustain the steady concentration of intracellular microcystin-LR. These findings dissect the response mechanism of M. aeruginosa to butachlor toxicity and provide valuable reference for the evaluation of potential risk caused by butachlor in aquatic environments.