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Freshwater resources are under stress around the world due to rapid urbanization and excessive water consumption. Cyanobacterial blooms have occurred frequently in surface waters, which produced toxic secondary metabolites causing a potential harm to aquatic ecosystems and humans. In this study, the relationship between different types of nitrogen source and the algal growth of Anabaena flos-aquae , which was isolated from Dianchi Lake in southern China, was investigated. Experiments were accomplished by using four types of isotope tracers including 15 N-ammonium chloride, 15 N-sodium nitrate, 15 N-urea, 15 N- l -alanine in culture medium to characterize the biosynthesis of 15 N-anatoxin-a (ATX-A), which is a major algal toxin from A. flos-aquae , through liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results showed that all these four types of nitrogen can be incorporated into algal cells. The ATX-A production with urea as the nitrogen source was much higher than that with the other three types of nitrogen. The 15 N labeling experiments further demonstrated that the uptake of organic nitrogen nutrients was significantly greater than that of inorganic nitrogen. These results provide new evidence and deeper insight to explore the biosynthesis of ATX-A in the specific strain of A. flos-aquae . [ABSTRACT FROM AUTHOR]

In order to identify the cyanobacterial species responsible of anatoxin-a (ATX) production in Lake Garda (Northern Italy), an intensive isolation and culturing of filamentous cyanobacteria were established since 2014 from environmental samples. In this work, we report a detailed account of the strategy adopted, which led to the discovery of a new unexpected producer of ATX, Tychonema bourrellyi . So far, this species is the first documented example of cultured Oscillatoriales able to produce ATX isolated from pelagic freshwater ecosystems. The isolated filaments were identified adopting a polyphasic approach, which included microscopic species identification, genetic characterisation and phylogenetic analyses based on 16S rRNA genes. The taxonomic identification was further confirmed by the high (>99%) rbcLX sequence similarities of the T. bourrellyi strains of Lake Garda with those deposited in DNA sequence databases. More than half of the isolates were shown to produce a significant amount of ATX, with cell quota ranging between 0.1 and 2.6 μg mm −3 , and 0.01 and 0.35 pg cell −1 . The toxic isolates were tested positive for anaC of the anatoxin-a synthetase ( ana ) gene cluster. These findings were confirmed with the discovery of one ATX producing T. bourrellyi strain isolated in Norway. This strain and a further non-ATX producing Norwegian Tychonema bornetii strain tested positive for the presence of the anaF gene of the ana gene cluster. Conversely, none of the Italian and Norwegian Tychonema strains were positive for microcystins (MCs), which was also confirmed by the absence of mcyE PCR products in all the samples analysed. This work suggests that the only reliable strategy to identify cyanotoxins producers should be based on the isolation of strains and their identification with a polyphasic approach associated to a concurrent metabolomic profiling. [ABSTRACT FROM AUTHOR]

Abstract: Two Advanced Oxidation Processes, namely vacuum-ultraviolet (VUV) photolysis at 172nm and ultraviolet/hydrogen peroxide (UV/H2O2) were investigated for the degradation of anatoxin-a in aqueous solutions. Solutions of anatoxin-a-fumarate were treated with VUV light at 172nm with a UV dose of 200 mJ/cm2, where fumaric acid served as a reference compound for a competition kinetics analysis. The second-order rate constant for the reaction between anatoxin-a and the hydroxyl radical was found to be (5.2±0.3)×109 M−1 s−1 and was independent of pH, temperature, and initial concentration of anatoxin-a. The direct photolysis of anatoxin-a using a medium pressure (MP) UV lamp was also investigated, in which case a UV dose of 1285 mJ/cm2 was required to degrade anatoxin-a by 88% and 50% at concentrations of 0.6mg/L and 1.8mg/L of toxin, respectively. Treatment of anatoxin-a with a low pressure (LP) UV lamp in the presence of 30mg/L of H2O2 was examined, where it was found that more than 70% of toxin could be degraded at a UV dose of 200 mJ/cm2. The degradation arises from the oxidation of the toxin by hydroxyl radicals. The addition of H2O2 clearly enhanced the degradation of anatoxin-a, up to a concentration of 40mg/L, after which addition of more H2O2 had little effect on the degradation kinetics of anatoxin-a. The effect of background constituents in the water on the degradation of anatoxin-a was also investigated using natural and synthetically produced model waters. [Copyright &y& Elsevier]

Cyanobacteria produce toxins that may contaminate drinking water sources. Among others, the presence of the alkaloid toxins cylindrospermopsin (CYN) and anatoxin-a (ANTX) constitutes a considerable threat to human health due to the acute and chronic toxicity of these compounds. In the present study, not previously reported second-order rate constants for the reactions of CYN and ANTX with chlorine and monochloramine and of CYN with potassium permanganate were determined and the influence of pH and temperature was established for the most reactive cases. It was found that the reactivity of CYN with chlorine presents a maximum at pH 7 (rate constant of 1265M−1 s−1). However, the oxidation of CYN with chloramine and permanganate are rather slow processes, with rate constants <1M−1 s−1. The first chlorination product of CYN was found to be 5-chloro-CYN (5-Cl-CYN), which reacts with chlorine 10–20 times slower than the parent compound. The reactivity of ANTX with chlorine and chloramines is also very low (k<1M−1 s−1). The elimination of CYN and ANTX in surface water was also investigated. A chlorine dose of 1.5mgl−1 was enough to oxidize CYN almost completely. However, 3mgl−1 of chlorine was able to remove only 8% of ANTX, leading to a total formation of trihalomethanes (TTHM) at a concentration of 150μgl−1. Therefore, chlorination is a feasible option for CYN degradation during oxidation and disinfection processes but not for ANTX removal. The permanganate dose required for CYN oxidation is very high and not applicable in waterworks. [Copyright &y& Elsevier]

Abstract: This study investigates the influence of chemical feed characteristics on nanofiltration performance for cyanotoxins removal, namely the neurotoxic anatoxin-a (alkaloid of 166g/mol, positively charged) and the hepatotoxic microcystins (cyclic peptides of ∼1000g/mol, negatively charged). Results indicate that NF membranes are an effective barrier against anatoxin-a and microcystins in drinking water. Anatoxin-a and especially microcystins were almost completely removed, regardless of the variations in feed water quality (natural organic matter and competitive toxin), the water recovery rate and the pH values. Anatoxin-a removal was governed by electrostatic interactions and steric hindrance, whereas for microcystins the latter was the main mechanism. In turn, fluxes were significantly impacted by background organics and, especially, inorganics (pH, calcium). [Copyright &y& Elsevier]

• Pico-cyanobacteria blooms driven by climate change are an emerging threat to water quality. • Cyanotoxin-producing pico-cyanobacteria were identified in tropical watersheds. • Cylindrospermopsin and anatoxin were detected in Synechococcus cultures. • Gene analogues for cylindrospermopsin production were discovered in Synechococcus. • Synechococcus may possess novel cylindrospermopsin synthesis pathways. Picocyanobacteria are small cyanobacteria, being about 0.8–1.5 µm in size. They are present in freshwater environments all over the world and are known to cause harmful algal blooms, although their effects are not well understood. Algal blooms are important to manage because they threaten freshwater resources, with potentially severe effects on ecological and human health. There is also increased urgency due to urbanization and climate change trends which are expected to exacerbate these bloom dynamics. These changes are expected to especially favour picocyanobacteria groups, emphasizing the need for better characterization of their effects in the environment. In this study, we report the discovery that Synechococcus sp. could produce cylindrospermopsin (CYN) and anatoxin-a (ATX). This ability had never been previously reported for this species. Their toxin genes were also partial compared to other major producers such as Raphidiopsis sp. and Anabaena sp., demonstrating potentially unique synthesis pathways that provides insight into the various mechanisms of genetic variation that drives toxin synthesis. The Synechococcus sp. strains were found to produce about 9.0 × 10−5–6.8 × 10−4 fg CYN cell−1 and 4.7 × 10−4–1.5 × 10−2 fg ATX cell−1. The potential for Synechococcus sp. to be toxic highlights a global concern due to its widespread distribution, and through environmental trends that increasingly favour its productivity within freshwater systems around the world. Novel cyanotoxin-producing Synechococcusspp. likely to become increasingly dominant in freshwater systems around the world, posing concerns that need to be better understood Image, graphical abstract [ABSTRACT FROM AUTHOR]

The occurrence of harmful cyanobacterial blooms has unabated increased over the last few decades, posing a significant risk for public health. In this work, we investigate the feasibility of catalytic wet peroxide oxidation (CWPO) promoted by modified natural magnetite (Fe 3 O 4 -R400/H 2 O 2), as an inexpensive, simple-operation and environmentally-friendly process for the removal of the cyanotoxins that show the major impact on drinking water: microcystins (MC-LR and MC-RR), cylindrospermopsin (CYN), anatoxin- a (ATX) and saxitoxin (STX). The performance of the system was evaluated under ambient conditions and circumneutral pH (pH 0 = 5) using relevant cyanotoxin concentrations (100–500 μg L−1). The nature of the cyanotoxins determined their reactivity towards CWPO, which decreased in the following order: MC-RR > CYN > MC-LR ≫ ATX > STX. In this sense, microcystins and CYN were completely removed in short reaction times (1–1.5 h) with a low catalyst concentration (0.2 g L−1) and the stoichiometric amount of H 2 O 2 (2–2.6 mg L−1), while only 60–80% conversion was achieved with ATX and STX in 5 h. In these cases, an intensification of the operating conditions (1 g L−1 catalyst and up to 30 mg H 2 O 2 L−1) was required to remove both toxins in 1 h. The impact of the main components of freshwaters i.e. natural organic matter (NOM) and several inorganic ions (HCO 3 −, HPO 4 2-, SO 4 2-) on the performance of the process was also investigated. Although the former led to a partial inhibition of the reaction due to HO· scavenging and catalyst coating, the latter did not show any remarkably effect, and the versatility of the process was finally confirmed in a real surface water. To further demonstrate the effectiveness of the catalytic system, the toxicity of both the initial cyanotoxins and the resulting CWPO effluents was measured with the brine shrimp Artemia salina. Remarkably, all CWPO effluents were non-toxic at the end of the treatment. Image 1 • The catalytic system Fe 3 O 4 -R400/H 2 O 2 is effective for the removal of cyanotoxins. • The reactivity of cyanotoxins decreased in the order: MC-RR > CYN > MC-LR ≫ ATX > STX. • The versatility of the process was demonstrated in different water matrices. • Oxidation effluents were non-toxic to the brine shrimp Artemia salina. [ABSTRACT FROM AUTHOR]