Spatiotemporal patterns and drivers of hepatotoxic microcystins (MC) were investigated in the Atchafalaya-Vermilion Bay System (AVBS), a subtropical, river-dominated estuary in Louisiana. Along with environmental data, monthly particulate MC (pMC) samples were examined over a two-year period (2016-2018), and biweekly pMC and dissolved MC (dMC) samples were examined over a five-month period in 2020. Solid phase adsorption toxin tracking (SPATT) samplers used to quantify time-integrated dMC concentrations were also deployed in 2020. Low, but detectable concentrations of pMC (≤0.033 μg L-1) and dMC (≤0.190 μg L-1) were found throughout the AVBS in 37.8 and 21.2 % of samples, respectively. Time integrative SPATT samplers detected dMC in nearly 100 % of the deployments, compared to dMC detections in 30.8 % of the discrete samples. This study documents widespread MC presence throughout the AVBS and while concentrations were low, knowledge gaps remain regarding the potential long-term impacts of sublethal MC exposure to estuarine organisms.

Harmful algal blooms and the toxins produced during these events are a human and environmental health concern worldwide. Saxitoxin and its derivatives are potent natural aquatic neurotoxins produced by certain freshwater cyanobacteria and marine algae species during these bloom events. Saxitoxins effects on human health are well studied, however its effects on aquatic biota are still largely unexplored. This work aims at evaluating the effects of a pulse acute exposure (24 h) of the model cladoceran Daphnia magna to 30 μg saxitoxin L-1, which corresponds to the safety guideline established by the World Health Organization (WHO) for these toxins in recreational freshwaters. Saxitoxin effects were assessed through a comprehensive array of biochemical (antioxidant enzymes activity and lipid peroxidation), genotoxicity (alkaline comet assay), neurotoxicity (total cholinesterases activity), behavioral (swimming patterns), physiological (feeding rate and heart rate), and epigenetic (total 5-mC DNA methylation) biomarkers. Exposure resulted in decreased feeding rate, heart rate, total cholinesterases activity and catalase activity. Contrarily, other antioxidant enzymes, namely glutathione-S-transferases and selenium-dependent Glutathione peroxidase had their activity increased, together with lipid peroxidation levels. The enhancement of the antioxidant enzymes was not sufficient to prevent oxidative damage, as underpinned by lipid peroxidation enhancement. Accordingly, average DNA damage level was significantly increased in STX-exposed daphnids. Total DNA 5-mC level was significantly decreased in exposed organisms. Results showed that even a short-term exposure to saxitoxin causes significant effects on critical molecular and cellular pathways and modulates swimming patterns in D. magna individuals. This study highlights sub-lethal effects caused by saxitoxin in D. magna, suggesting that these toxins may represent a marked challenge to their thriving even at a concentration deemed safe for humans by the WHO.

Microcystins (MCs) can be detected in various matrices in two forms: a freely extractable fraction and a total (free and covalently protein-bound) fraction. Although the majority of MCs analyses are limited to the free fraction, they do not allow the analysis of all MCs variants or protein-bound forms. Other methods, known as total MCs analysis methods, enable simultaneous analysis of all MCs variants, as well as bound forms, which may be a major form of toxin accumulation in organisms. Among these techniques, the chemical oxidation method (e.g. Lemieux) allows the detection of total forms of MC (and nodularins) by oxidizing the common part to all MC and nodularins, and analyzing the resultant MMPB product (2-methyl-3-methoxy-4-phenylbutyric acid). However, the execution of this method in the context of health monitoring is challenging due to the variability of the protocols, the recoveries obtained with these protocols, and the important matrix effects associated with the method. The objectives of this study were i) to optimize an existing protocol of chemical oxidation \"Lemieux1\" on fresh fish fillet matrices, ii) to compare two existing protocols (\"Lemieux1\" and \"Lemieux2\"), and iii) apply Lemieux oxidation to fish fillets and livers naturally contaminated with MCs-producing cyanobacteria and to freshwater mussels contaminated with MCs in laboratories. Optimization of the \"Lemieux1\" protocol, in particular in the oxidation and SPE (solid phase extraction) steps improved the method\'s yields on the fresh fish fillet matrix (from <5 % to around 40 %). Moreover, several quantification methods have been compared through various calibration techniques (solvent calibration curve, matrix-matched calibration curve, oxidized MC-LR calibration curve and also by testing the addition of d3-MMPB as an internal standard). Comparison with the \"Lemieux2\" protocol showed the best results on the same matrix, with yields of around 65 %. MMPB was analyzed using this \"Lemieux 2\" protocol, in livers of carps sampled during an episode of cyanobacteria proliferation, at concentrations ranging from 17.9 to 27.5 μg/kg MMPB and at concentrations ranging from 50 to 2890 μg/kg MMPB in freshwater mussels laboratory contaminated to MCs.

Cyanobacteria in water supplies are considered an emerging threat, as some species produce toxic metabolites, cyanotoxins, of which the most widespread and well-studied are microcystins. Consumption of contaminated water is a common exposure route to cyanotoxins, making the study of cyanobacteria in drinking waters a priority to protect public health. In drinking water treatment plants, pre-oxidation with chlorinated compounds is widely employed to inhibit cyanobacterial growth, although concerns on its efficacy in reducing cyanotoxin content exists. Additionally, the effects of chlorination on abundant but less-studied cyanometabolites (e.g. cyanopeptolins whose toxicity is still unclear) remain poorly investigated. Here, two chlorinated oxidants, sodium hypochlorite (NaClO) and chlorine dioxide (ClO2), were tested on the toxic cyanobacterium Microcystis aeruginosa, evaluating their effect on cell viability, toxin profile and content. Intra- and extracellular microcystins and other cyanometabolites, including their degradation products, were identified using an untargeted LC-HRMS approach. Both oxidants were able to inactivate M. aeruginosa cells at a low dose (0.5 mg L-1), and greatly reduced intracellular toxins content (>90%), regardless of the treatment time (1-3 h). Conversely, a two-fold increase of extracellular toxins after NaClO treatment emerged, suggesting a cellular damage. A novel metabolite named cyanopeptolin-type peptide-1029, was identified based on LC-HRMSn (n = 2, 3) evidence, and it was differently affected by the two oxidants. NaClO led to increase its extracellular concentration from 2 to 80-100 μg L-1, and ClO2 induced the formation of its oxidized derivative, cyanopeptolin-type peptide-1045. In conclusion, pre-oxidation treatments of raw water contaminated by toxic cyanobacteria may lead to increased cyanotoxin concentrations in drinking water and, depending on the chemical agent, its dose and treatment duration, also of oxidized metabolites. Since the effects of such metabolites on human health remain unknown, this issue should be handled with extreme caution by water security agencies involved in drinking water management.

Harmful cyanobacterial blooms will be more intense and frequent in the future, contaminating surface waters with cyanotoxins and posing a threat to communities heavily reliant on surface water usage for crop irrigation. Constructed wetlands (CWs) are proposed to ensure safe crop irrigation, but more research is needed before implementation. The present study operated 28 mesocosms in continuous mode mimicking horizontal sub-surface flow CWs. Mesocosms were fed with synthetic lake water and spiked periodically with two cyanotoxins, microcystin-LR (MC-LR) and cylindrospermopsin (CYN), at environmentally relevant cyanotoxins concentrations (10 μg L-1). The influence of various design factors, including plant species, porous media, and seasonality, was explored. The mesocosms achieved maximum MC-LR and CYN mass removal rates of 95 % and 98 %, respectively. CYN removal is reported for the first time in CWs mimicking horizontal sub-surface flow CWs. Planted mesocosms consistently outperformed unplanted mesocosms, with Phragmites australis exhibiting superior cyanotoxin mass removal compared to Juncus effusus. Considering evapotranspiration, J. effusus yielded the least cyanotoxin-concentrated effluent due to the lower water losses in comparison with P. australis. Using the P-kC* model, different scaling-up scenarios for future piloting were calculated and discussed. Additionally, bacterial community structure was analyzed through correlation matrices and differential taxa analyses, offering valuable insights into their removal of cyanotoxins. Nevertheless, attempts to validate microcystin-LR biotransformation via the known mlrA gene degradation pathway were unfruitful, indicating alternative enzymatic degradation pathways occurring in such complex CW systems. Further investigation into the precise molecular mechanisms of removal and the identification of transformation products is needed for the comprehensive understanding of cyanotoxin mitigation in CW. This study points towards the feasibility of horizontal sub-surface flow CWs to be employed to control cyanotoxins in irrigation or recreational waters.

Algal blooms can threaten human health if cyanotoxins such as microcystin are produced by cyanobacteria. Regularly monitoring microcystin concentrations in recreational waters to inform management action is a tool for protecting public health; however, monitoring cyanotoxins is resource- and time-intensive. Statistical models that identify waterbodies likely to produce microcystin can help guide monitoring efforts, but variability in bloom severity and cyanotoxin production among lakes and years makes prediction challenging. We evaluated the skill of a statistical classification model developed from water quality surveys in one season with low temporal replication but broad spatial coverage to predict if microcystin is likely to be detected in a lake in subsequent years. We used summertime monitoring data from 128 lakes in Iowa (USA) sampled between 2017 and 2021 to build and evaluate a predictive model of microcystin detection as a function of lake physical and chemical attributes, watershed characteristics, zooplankton abundance, and weather. The model built from 2017 data identified pH, total nutrient concentrations, and ecogeographic variables as the best predictors of microcystin detection in this population of lakes. We then applied the 2017 classification model to data collected in subsequent years and found that model skill declined but remained effective at predicting microcystin detection (area under the curve, AUC ≥ 0.7). We assessed if classification skill could be improved by assimilating the previous years\' monitoring data into the model, but model skill was only minimally enhanced. Overall, the classification model remained reliable under varying climatic conditions. Finally, we tested if early season observations could be combined with a trained model to provide early warning for late summer microcystin detection, but model skill was low in all years and below the AUC threshold for two years. The results of these modeling exercises support the application of correlative analyses built on single-season sampling data to monitoring decision-making, but similar investigations are needed in other regions to build further evidence for this approach in management application.

Anatoxin-a (ATX-a) is a neurotoxin produced by some species of cyanobacteria. Due to its water solubility and stability in natural water, it could pose health risks to human, animals, and plants. Conventional water treatment techniques are not only insufficient for the removal of ATX-a, but they also result in cell lysis and toxin release. The elimination of this toxin through biodegradation may be a promising strategy. This study examines for the first time the biodegradation of ATX-a to a non-toxic metabolite (Epoxy-ATX-a) by a strain of Bacillus that has a history of dealing with toxic cyanobacteria in a eutrophic lake. The Bacillus strain AMRI-03 thrived without lag phase in a lake water containing ATX-a. The strain displayed fast degradation of ATX-a, depending on initial toxin concentration. At the highest initial concentrations (50 & 100 µg L- 1), total ATX-a degradation took place in 4 days, but it took 6 & 7 days at lower concentrations (20, 10, and 1 µg L- 1, respectively). The ATX-a biodegradation rate was also influenced by the initial toxin concentration, reaching its maximum value (12.5 µg L- 1 day- 1) at the highest initial toxin concentrations (50 & 100 µg L- 1). Temperature and pH also had an impact on the rate of ATX-a biodegradation, with the highest rates occurring at 25 and 30 ºC and pH 7 and 8. This nontoxic bacterial strain could be immobilized within a biofilm on sand filters and/or sludge for the degradation and removal of ATX-a and other cyanotoxins during water treatment processes, following the establishment of mesocosm experiments to assess the potential effects of this bacterium on water quality.

Biocrusts dominate the soil surface in deserts and are composed of diverse microbial communities that provide important ecosystem services. Cyanobacteria in biocrusts produce many secondary metabolites, including the neurotoxins BMAA, AEG, DAB, anatoxin-a(S) (guanitoxin), and the microcystin hepatotoxins, all known or suspected to cause disease or illness in humans and other animals. We examined cyanobacterial growth and prevalence of these toxins in biocrusts at millimeter-scales, under a desert-relevant illumination gradient. In contrast to previous work, we showed that hydration had an overall positive effect on growth and toxin accumulation, that nitrogen was not correlated with growth or toxin production, and that phosphorus enrichment negatively affected AEG and BMAA concentrations. Excess illumination positively correlated with AEG, and negatively correlated with all other toxins and growth. Basic pH negatively affected only the accumulation of BMAA. Anatoxin-a(S) (guanitoxin) was not correlated with any tested variables, while microcystins were not detected in any of the samples. Concerning toxin pools, AEG and BMAA were good predictors of the presence of one another. In a newly conceptualized scheme, we integrate aspects of biocrust growth and toxin pool accumulations with arid-relevant desertification drivers.

Harmful algal blooms (HABs) are a persistent and increasing problem globally, yet we still have limited knowledge about how they affect wildlife. Although semi-aquatic and aquatic amphibians and reptiles have experienced large declines and occupy environments where HABs are increasingly problematic, their vulnerability to HABs remains unclear. To inform monitoring, management, and future research, we conducted a literature review, synthesized the studies, and report on the mortality events describing effects of cyanotoxins from HABs on freshwater herpetofauna. Our review identified 37 unique studies and 71 endpoints (no-observed-effect and lowest-observed-effect concentrations) involving 11 amphibian and 3 reptile species worldwide. Responses varied widely among studies, species, and exposure concentrations used in experiments. Concentrations causing lethal and sublethal effects in laboratory experiments were generally 1 to 100 µg/L, which contains the mean value of reported HAB events but is 70 times less than the maximum cyanotoxin concentrations reported in the environment. However, one species of amphibian was tolerant to concentrations of 10,000 µg/L, demonstrating potentially immense differences in sensitivities. Most studies focused on microcystin-LR (MC-LR), which can increase systemic inflammation and harm the digestive system, reproductive organs, liver, kidneys, and development. The few studies on other cyanotoxins illustrated that effects resembled those of MC-LR at similar concentrations, but more research is needed to describe effects of other cyanotoxins and mixtures of cyanotoxins that commonly occur in the environment. All experimental studies were on larval and adult amphibians; there were no such studies on reptiles. Experimental work with reptiles and adult amphibians is needed to clarify thresholds of tolerance. Only nine mortality events were reported, mostly for reptiles. Given that amphibians likely decay faster than reptiles, which have tissues that resist decomposition, mass amphibian mortality events from HABs have likely been under-reported. We propose that future efforts should be focused on seven major areas, to enhance our understanding of effects and monitoring of HABs on herpetofauna that fill important roles in freshwater and terrestrial environments. Environ Toxicol Chem 2024;00:1-14. Published 2024. This article is a U.S. Government work and is in the public domain in the USA. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Microcystin (MC), a hepatotoxin that is harmful to human health, has frequently increased in freshwaters worldwide due to the increase in toxic cyanobacterial blooms. Despite many studies reported the human exposure to MC through drinking water, the potential transfer of this toxin to human via consumption of vegetables grown on farmlands that are naturally irrigated with contaminated water has not been largely investigated. Therefore, this study investigates the presence of MC in irrigation water and its potential accumulation in commonly consumed vegetables from Egyptian farmlands. The results of toxin analysis revealed that all irrigation water sites contained high MC concentrations (1.3-93.7 μg L-1) along the study period, in association with the abundance of dominant cyanobacteria in these sites. Meanwhile, MCs were detected in most vegetable plants surveyed, with highest levels in potato tubers (1100 μg kg-1 fresh weight, FW) followed by spinach (180 μg kg-1 FW), onion (170 μg g-1 FW), Swiss chard (160 μg kg-1 FW) and fava bean (46 μg kg-1 FW). These MC concentrations in vegetables led to estimated daily intake (EDI) values (0.08-1.13 μg kg bw-1 d-1 for adults and 0.11-1.5 μg kg bw-1 d-1 for children), through food consumption, exceeding the WHO recommended TDI (0.04 μg kg bw-1 d-1) for this toxin. As eutrophic water is widely used for irrigation in many parts of the world, our study suggests that cyanotoxins in irrigation waters and agricultural plants should be regularly monitored to safeguard the general public from inadvertent exposure to harmful toxins via food consumption.