Concentrations Of microcystin-LR Research Articles

Microcystin-LR Induces Estrogenic Effects at Environmentally Relevant Concentration in Black-Spotted Pond Frogs (Pelophylax nigromaculatus): In Situ, In Vivo, In Vitro, and In Silico Investigations.

Harmful cyanobacterial blooms are frequent and intense worldwide, creating hazards for aquatic biodiversity. The potential estrogen-like effect of Microcystin-LR (MC-LR) is a growing concern. In this study, we assessed the estrogenic potency of MC-LR in black-spotted frogs through combined field and laboratory approaches. In 13 bloom areas of Zhejiang province, China, the MC-LR concentrations in water ranged from 0.87 to 8.77 μg/L and were correlated with sex hormone profiles in frogs, suggesting possible estrogenic activity of MC-LR. Tadpoles exposed to 1 μg/L, an environmentally relevant concentration, displayed a female-biased sex ratio relative to controls. Transcriptomic results revealed that MC-LR induces numerous and complex effects on gene expression across multiple endocrine axes. In addition, exposure of male adults significantly increased the estradiol (E2)/testosterone (T) ratio by 3.5-fold relative to controls. Downregulation of genes related to male reproductive endocrine function was also identified. We also showed how MC-LR enhances the expression of specific estrogen receptor (ER) proteins, which induce estrogenic effects by activating the ER pathway and hypothalamic-pituitary-gonadal (HPG) axis. In aggregate, our results reveal multiple lines of evidence demonstrating that, for amphibians, MC-LR is an estrogenic endocrine disruptor at environmentally relevant concentrations. The data presented here support the need for a shift in the MC-LR risk assessment. While hepatoxicity has historically been the focus of MC-LR risk assessments, our data clearly demonstrate that estrogenicity is a major mode of toxicity at environmental levels and that estrogenic effects should be considered for risk assessments on MC-LR going forward.

Control of Microcystis aeruginosa by Daphnia: Experimental evidence and identification of involved infochemicals.

Infochemicals refer to chemicals responsible for information exchange between organisms. We evaluated the effects of Daphnia magna and Daphnia galeata infochemicals on Microcystis aeruginosa for 15d. The Daphnia infochemicals were obtained from spent medium after culturing Daphnia in Elendt M4 medium for 48 h. Both Daphnia infochemicals significantly increased (p < 0.05) the intracellular reactive oxygen species level and microcystin-LR concentration in M. aeruginosa. This cellular effect increased colony formation of M. aeruginosa, thereby inhibiting the growth of M. aeruginosa. D. galeata infochemicals provoked significantly greater (p < 0.05) adverse effects on M. aeruginosa than those of D. magna infochemicals, which were further exaggerated by pre-exposure of Daphnia to M. aeruginosa. This result seems to be related to the different compositions and concentrations of Daphnia infochemicals. Several Daphnia infochemicals, such as methyl ferulate, cyclohexanone, 3, 5-dimethyl, hexanedioic acid, and bis(2-ethylhexyl) ester, showed a high correlation with M. aeruginosa cell concentration (|r | >0.6), suggesting that they may play a key role in controlling harmful cyanobacteria. Additionally, pre-exposure of D. magna and D. galeata to M. aeruginosa produced oleic acid, methyl ester, and n-hexadecanoic acid, with a highly correlation with M. aeruginosa cell concentration (|r | >0.6). p-tolyl acetate and linoleic acid were detected only in the pre-exposed D. galeata infochemicals. These findings suggest that some of Daphnia infochemicals identified in this study can be a promising tool to control M. aeruginosa growth. However, further studies are required to verify the specific actions of these infochemicals against cyanobacteria.

Magnetic recyclable visible light-driven Bi2WO6/Fe3O4/RGO for photocatalytic degradation of Microcystin-LR: Mechanism, pathway, and influencing factors

Photocatalysis was an attractive strategy that had potential to tackle the Microcystin-LR (MC-LR) contamination of aquatic ecosystems. Herein, magnetic photocatalyst Fe3O4/Bi2WO6/Reduced graphene oxide composites (Bi2WO6/Fe3O4/RGO) were employed to degrade MC-LR. The removal efficiency and kinetic constant of the optimized Bi2WO6/Fe3O4/RGO (Bi2WO6/Fe3O4-40%/RGO) was 1.8 and 2.3 times stronger than the pure Bi2WO6. The improved activity of Bi2WO6/Fe3O4-40%/RGO was corresponded to the expanded visible light adsorption ability and reduction of photogenerated carrier recombination efficiency through the integration of Bi2WO6 and Fe3O4-40%/RGO. The MC-LR removal efficiency exhibited a positive tendency to the initial density of algae cells, fulvic acid, and the concentration of MC-LR decreased. The existed anions (Cl−, CO3−2, NO3−, H2PO4−) reduced MC-LR removal efficiency of Bi2WO6/Fe3O4-40%/RGO. The Bi2WO6/Fe3O4-40%/RGO could degrade 79.3% of MC-LR at pH = 7 after 180 min reaction process. The trapping experiments and ESR tests confirmed that the h+, ∙OH, and ∙O2− played a significant role in MC-LR degradation. The LC-MS/MS result revealed the intermediates and possible degradation pathways.

Changes in physiology, antioxidant system, and gene expression in Microcystis aeruginosa under fenoxaprop-p-ethyl stress.

Fenoxaprop-p-ethyl (FE) is one of the typical aryloxyphenoxypropionate herbicides. FE has been widely applied in agriculture in recent years. Human health and aquatic ecosystems are threatened by the cyanobacteria blooms caused by Microcystis aeruginosa, which is one of the most common cyanobacteria responsible for freshwater blooming. Few studies have been reported on the physiological effects of FE on M. aeruginosa. This study analyzed the growth curves, the contents of chlorophyll a and protein, the oxidative stress, and the microcystin-LR (MC-LR) levels of M. aeruginosa exposed to various FE concentrations (i.e., 0, 0.5, 1, 2, and 5mg/L). FE was observed to stimulate the cell density, chlorophyll a content, and protein content of M. aeruginosa at 0.5- and 1-mg/L FE concentrations but inhibit them at 2 and 5mg/L FE concentrations. The superoxide dismutase and catalase activities were enhanced and the malondialdehyde concentration was increased by FE. The intracellular (intra-) and extracellular (extra-) MC-LR contents were also affected by FE. The expression levels of photosynthesis-related genes psbD1, psaB, and rbcL varied in response to FE exposure. Moreover, the expressions of microcystin synthase-related genes mcyA and mcyD and microcystin transportation-related gene mcyH were significantly inhibited by the treatment with 2 and 5mg/L FE concentrations. These results might be helpful in evaluating the ecotoxicity of FE and guiding the rational application of herbicides in modern agriculture.

Experimental accumulation and depuration kinetics and natural occurrence of microcystin-LR in basil (Ocimum basilicum L.)

Microcystin-LR (MC-LR) is a hepatotoxic metabolite that naturally occurs during some cyanobacterial blooms in eutrophic waterbodies, and irrigation of edible plants with MC-LR-contaminated water causes bioaccumulation of the toxin. However, sufficient information about accumulation and depuration mechanics in hydroculture-grown herb plants is still lacking. This work aimed at 1) investigating bioaccumulation and depuration of MC-LR in basil, 2) verifying the possible MC-LR detoxification mechanisms in the plant, and 3) detecting the natural occurrence of MC-LR in basil (n = 50) collected from the Belgian market. Basil plants grown in a hydroculture were exposed to MC-LR (5, 20, and 50 μg L−1) spiked in a Hoagland solution for seven days. MC-LR depuration was also studied by transferring the plants to a non-contaminated Hoagland solution after exposure to MC-LR for another seven days. MC-LR concentrations in Hoagland solution, basil leaves, and roots were quantified using a validated UHPLC–MS/MS method. In addition, ELISA and LC–HRMS (only basil leaves) were used for confirmation. The results showed an increase in the accumulated levels of MC-LR at higher exposure doses, with higher MC-LR levels in roots than in leaves for all the treatment conditions. For MC-LR depuration, significant reductions were observed in all the treatment conditions for roots only. No MC-LR conjugates, potentially related to metabolism, were detected by LC–HRMS. Finally, MC-LR was detected in one store-bought basil sample, representing the first occurrence of cyanotoxins in an edible crop from Belgium.

Degradation performance and mechanism of microcystins in aquaculture water using low-temperature plasma technology

The eutrophication of aquaculture water bodies seriously restricts the healthy development of the aquaculture industry. Among them, microcystins are particularly harmful. Therefore, the development of technologies for degrading microcystins is of great significance for maintaining the healthy development of the aquaculture industry. The feasibility and mechanism of removing microcystins-LR by dielectric barrier discharge (DBD) plasma were studied. DBD discharge power of 49.6 W and a treatment time of 40 min were selected as the more suitable DBD parameters, resulting in microcystin-LR removal efficiency of 90.4%. Meanwhile, the effects of initial microcystin-LR concentration, initial pH value, turbidity, anions on the degradation effect of microcystin-LR were investigated. The removal efficiency of microcystin-LR decreased with the increase of initial microcystin-LR concentration and turbidity. The degradation efficiency of microcystin-LR at pH 4.5 and 6.5 is significantly higher than that at pH 8.5 and 3.5. HCO3− can inhibit the removal efficiency of microcystin-LR. Furthermore, five intermediates products (m/z = 1029.5, 835.3, 829.3, 815.4, 642.1) were identified in this study, and the toxicity analysis of these degradation intermediates indicated that DBD treatment can reduce the toxicity of microcystin-LR. e－aq, •OH, H2O2, and O3 have been shown to play a major role in the degradation of microcystin-LR, and the contribution ranking of these active species is e－aq > •OH > H2O2 > O3. The application of DBD plasma technology in microcystin-LR removal and detoxification has certain development potential.

Effects of the catastrophic 2020 Yangtze River seasonal floods on microcystins and environmental conditions in Three Gorges Reservoir Area, China.

During July and August 2020, Three Gorges Reservoir Area (TGRA) suffered from catastrophic seasonal floods. Floods changed environmental conditions and caused increase in concentration of microcystins (MCs) which is a common and potent cyanotoxin. However, the effects and seasonal variations of MCs, cyanobacteria, and environmental conditions in TGRA after the 2020 Yangtze River extreme seasonal floods remain largely unclear, and relevant studies are lacking in the literature. A total of 12 representative sampling sites were selected to perform concentration measurement of relevant water quality objectives and MCs in the representative area of the TGRA. The sampling period was from July 2020 to October 2021, which included the flood period. Organic membrane filters were used to perform the DNA extraction and analyses of the 16S rRNA microbiome sequencing data. Results showed the seasonal floods result in significant increases in the mean values of microcystin-RR (MCRR), microcystin-YR (MCYR), and microcystin-LR (MCLR) concentration and some water quality objectives (i.e., turbidity) in the hinterland of TGRA compared with that in non-flood periods (p < 0.05). The mean values of some water quality objectives (i.e., total nitrogen (TN), total phosphorus (TP), total dissolved phosphorus (TDP), and turbidity), MC concentration (i.e., MCRR, MCYR, and MCLR), and cyanobacteria abundance (i.e., Cyanobium_PCC-6307 and Planktothrix_NIVA-CYA_15) displayed clear tendency of increasing in summer and autumn and decreasing in winter and spring in the hinterland of TGRA. The results suggest that seasonal floods lead to changes in MC concentration and environmental conditions in the hinterland of TGRA. Moreover, the increase in temperature leads to changes in water quality objectives, which may cause water eutrophication. In turn, water eutrophication results in the increase in cyanobacteria abundance and MC concentration. In particular, the increased MC concentration may further contribute to adverse effects on human health.

Effects of microcystin-LR on purification efficiency of simulating drinking water source by Hydrocharis dubia (Bl.) backer

The safety of drinking water source directly affects human health. Microcystin-LR (MC-LR), a toxic and common pollutant in drinking water source, is released by algae and can impede the in-situ remediation effect of aquatic plant. Finding out the effect mechanism of MC-LR on the purification of drinking water by aquatic plant is the key to its application. This study aims to explore the performance and mechanism of MC-LR on drinking water source purification by Hydrocharis dubia (Bl.) backer. The optimum removal efficiency of NH4+-N, TP and COD were 90.7%, 93.2% and 77.3% at MC-LR concentration of 0.5 μg L−1, respectively. With the increase of MC-LR concentration, the pollutants removal rate was obviously inhibited causing by concentration-dependent. Furthermore, the growth and development of the Hydrocharis dubia (Bl.) backer roots were significantly promoted at the concentration of 0.1 μg L−1. The length, tips, surface area, and average diameter of the root increased by 71.3%, 271.4%, 265.5%, and 113.0%, respectively. Chlorophyll contents under low-concentration MC-LR show a 14.5%–15.7% promoting effect compared with the control group. The activities of POD and CAT were also stimulated with the MC-LR increasing (<1.0 μg L−1). Notably, the MDA contents increased with increasing MC-LR concentration (p < 0.01). This study indicates the effect mechanism of MC-LR on Hydrocharis dubia (Bl.) backer purification performance relies on the increased growth and enzyme activity of Hydrocharis dubia (Bl.) backer.

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.