Isolation and characterization of a novel Lambda-like phage infecting the bloom-forming cyanobacteria Cylindrospermopsis raciborskii.

Cylindrospermopsis raciborskii is a central bloom-forming cyanobacteria. However, despite its ecological significance, little is known of its interactions with the phages that infect it. Currently, only a single sequenced genome of a Cylindrospermopsis-infecting phage is publicly available. Here we describe the isolation and characterization of Cr-LKS3, a second phage infecting Cylindrospermopsis. Cr-LKS3 is a siphovirus with a higher genome similarity to prophages within heterotrophic bacteria genomes than to any other cyanophage/cyano-prophage, suggesting that it represents a novel cyanophage group. The function, order and orientation of the 72 genes in the Cr-LKS3 genome are highly similar to those of Escherichia virus Lambda (hereafter Lambda), despite the very low sequence similarity between these phages, showing high evolutionary convergence despite the substantial difference in host characteristics. Similarly to Lambda, the genome of Cr-LKS3 contains various genes that are known to be central to lysogeny, suggesting it can enter a lysogenic cycle. Cr-LKS3 has a unique ability to infect a host with a dramatically different GC content, without carrying any tRNA genes to compensate for this difference. This ability, together with its potential lysogenic lifestyle shed light on the complex interactions between C. raciborskii and its phages.

Comparative proteomics of the toxigenic diazotroph Raphidiopsis raciborskii (cyanobacteria) in response to iron.

Raphidiopsis raciborskii is an invasive bloom-forming cyanobacteria with the flexibility to utilize atmospheric and fixed nitrogen. Since nitrogen-fixation has a high requirement for iron as an ezyme cofactor, we hypothesize that iron availability would determine the success of the species under nitrogen-fixing conditions. This study compares the proteomic response of cylindrospermopsin-producing and non-toxic strains of R. racibroskii to reduced iron concentrations, under nitrogen-fixing conditions, to examine any strain-specific adaptations that might increase fitness under these conditions. We also compared their proteomic responses at exponential and stationary growth phases to capture the changes throughout the growth cycle. Overall, the toxic strain was more competitive under Fe-starved conditions during exponential phase, with upregulated growth and transport-related proteins. The non-toxic strain showed reduced protein expression across multiple primary metabolism pathways. We propose that the increased expression of porin proteins during the exponential growth phase enables toxic strains to persist under Fe-starved conditions with this ability providing a potential explanation for the increased fitness of cylindrospermoipsin-producing strains during unfavourable environmental conditions.

Heterologous Expression of an Unusual Ketosynthase, SxtA, Leads to Production of Saxitoxin Intermediates in Escherichia coli.

Paralytic shellfish toxins (PSTs) are neurotoxic alkaloids produced by freshwater cyanobacteria and marine dinoflagellates. Due to their antagonism of voltage-gated sodium channels in excitable cells, certain analogues are of significant pharmacological interest. The biosynthesis of the parent compound, saxitoxin, is initiated with the formation of 4-amino-3-oxo-guanidinoheptane (ethyl ketone) by an unusual polyketide synthase-like enzyme, SxtA. We have heterologously expressed SxtA from Raphidiopsis raciborskii T3 in Escherichia coli and analysed its activity in vivo. Ethyl ketone and a truncated analogue, methyl ketone, were detected by HPLC-ESI-HRMS analysis, thus suggesting that SxtA has relaxed substrate specificity in vivo. The chemical structures of these products were further verified by tandem mass spectrometry and labelled-precursor feeding with [guanidino-15 N2 ] arginine and [1,2-13 C2 ] acetate. These results indicate that the reactions catalysed by SxtA could give rise to multiple PST variants, including analogues of ecological and pharmacological significance.

Elevated co2 has Differential Effects on Five Species of Microalgae from a Subtropical Freshwater Lake: Possible Implications for Phytoplankton Species Composition.

Rising atmospheric CO2 concentrations are predicted to have a significant impact on global phytoplankton populations. Of particular interest in freshwater systems are those species that produce toxins or impact water quality, though evidence for how these species, and many others, will respond is limited. This study investigated the effects of elevated CO2 (1,000 ppm) relative to current atmospheric CO2 partial pressures (400 ppm), on growth, cell size, carbon acquisition, and photophysiology of five freshwater phytoplankton species including a toxic cyanophyte, Raphidiopsis raciborskii, from Lake Wivenhoe, Australia. Effects of elevated CO2 on growth rate varied between species; notably growth rate was considerably higher for Staurastrum sp. and significantly lower for Stichococcus sp. with a trend to lower growth rate for R. raciborskii. Surface area to volume ratio was significantly lower with elevated CO2 , for all species except Cyclotella sp. Timing of maximum cell concentrations of those genera studied in monoculture occurred in the lake in order of CO2 affinity when free CO2 concentrations dropped below air equilibrium. The results presented here suggest that as atmospheric levels of CO2 rise, R. raciborskii may become less of a problem to water quality, while some species of chlorophytes may become more dominant. This has implications for stakeholders of many freshwater systems.

Inhibition of growth rate and cylindrospermopsin synthesis by Raphidiopsis raciborskii upon exposure to macrophyte Lemna trisulca (L).

Impact of macrophyte Lemna trisulca on the growth rate and synthesis of cylindrospermopsin (CYN) by cyanobacterium Raphidiopsis raciborskii was determined. The presence of L. trisulca inhibited the biomass accumulation of the cyanobacterium by 25% compared to the control during co-cultivation. The simultaneous cultivation of these organisms slightly affected the inhibition of macrophyte growth rate by 5.5% compared to the control. However, no morphological changes of L. trisulca after incubation with cyanobacteria were observed. It was also shown that the long-term (35 days) co-cultivation of R. raciborskii and L. trisulca led to a decrease in CYN concentration in media and cyanobacterial cells by 32 and 38%, respectively, compared to the values obtained for independent cultivation of cyanobacterium. Excessive absorption of phosphate ions by L. trisulca from the medium compared to nitrate ions led to a significant increase in the nitrate:phosphate ratio in the media, which inhibits the development of cyanobacterium. The obtained results indicate that L. trisulca in the natural environment may affect the physiology of cyanobacteria. The presented study is the first assessment of the allelopathic interaction of macrophyte and R. raciborskii.

Interactive effect of water level and flushing rate on population dynamics of a harmful cyanobacterial species: Raphidiopsis raciborskii.

Changes in water level and flushing rate directly affect to a large extent the biomass of harmful cyanobacteria, and drive the shift of phytoplankton composition between cyanobacteria dominance/non-dominance in eutrophic waters. Here, we gave a theoretical formula describing the combinational effect of water level and flushing rate on cyanobacterial biomass in eutrophic and well-mixed waters. We also formulated an equation predicting the water level and flushing rate at which cyanobacteria become non-dominating in such water columns. The formulae were confronted with field observations of a low-light adapted cyanobacterium in a large coastal reservoir of southern China. Our formulae demonstrate that water level and flushing rate have an interactive effect on the equilibrium biomass of low-light adapted cyanobacteria in mixed and turbid waters. The formulae were well fitted to the field observation of Raphidiopsis raciborskii population in the reservoir during four dry seasons. In agreement with the theoretical analysis, multiple regression analysis also showed that the interaction between water level and flushing rate is able to interpret the variation of R. raciborskii biomass in the water column. The two formulae are applicable for predicting the response of low-light adapted cyanobacteria to local climate change. Our findings have practical significance in designing measures against the dominance of low light-adapted cyanobacteria in reservoirs.

Evidence of cylindrospermopsin uptake and clearance in fish (Oreochromis niloticus) under laboratory conditions.

Cylindrospermopsin (CYN) is a cyanotoxin that has raised serious concerns about public health in many parts of the world. It can bioaccumulate and affect the health of aquatic organisms, but despite this, few studies have been conducted on CYN uptake and clearance in fish. In this paper, the authors evaluate the uptake and clearance of CYN in the muscle tissue and viscera of juvenile tilapia (Oreochromis niloticus) after exposure to aqueous extracts and whole cells of Cylindrospermopsis raciborskii (CYN-producer). CYN blended with commercial fish food, and three experiments were conducted. In the first trial, fish food, and aqueous extracts containing 0.31 µg CYN g-1 of food per day, was administered to tilapia for 15 days. In the second trial, fish were provided food and intact cells (5.4 µg CYN g-1 of food per day) for 15 days. In the last trial, they were provided fish food and aqueous extracts (0.8 µg CYN g-1 of food per day) for 12 days, and for the next 10 days, the animals were fed food without toxic cell extracts (to simulate a clearance period). The concentration of CYN in muscle tissue and viscera was analysed using ELISA. In the case of juvenile tilapia, the presence of CYN was higher in viscera than in muscle tissue, and the toxin remained in the tissues even after 10 days without the addition of contaminated food. The results suggest that tilapia represents a potential source of CYN transfer through the food web, and this shows the need for a continuous monitoring of this compound in organisms that are used for human and animal consumption.

Biogeography of the cyanobacterium Raphidiopsis (Cylindrospermopsis) raciborskii: Integrating genomics, phylogenetic and toxicity data.

Raphidiopsis (Cylindrospermopsis) raciborskii, a globally distributed bloom-forming cyanobacterium, produces either the cytotoxin cylindrospermopsin (CYL) in Oceania, Asia and Europe or the neurotoxin saxitoxin (STX) and analogues (paralytic shellfish poison, PSP) in South America (encoded by sxt genetic cluster) and none of them in Africa. Nevertheless, this particular geographic pattern is usually overlooked in current hypotheses about the species dispersal routes. Here, we combined genomics, phylogenetic analyses, toxicity data and a literature survey to unveil the evolutionary history and spread of the species. Phylogenies based on 354 orthologous genes from all the available genomes and ribosomal ITS sequences of the taxon showed two well-defined clades: the American, having the PSP producers; and the Oceania/Europe/Asia, including the CYL producers. We propose central Africa as the original dispersion center (non-toxic populations), reaching North Africa and North America (in former Laurasia continent). The ability to produce CYL probably took place in populations that advanced to sub-Saharan Africa and then to Oceania and South America. According to the genomic context of the sxt cluster found in PSP-producer strains, this trait was acquired once by horizontal transfer in South America, where the ability to produce CYL was lost.

Effects and possible mechanisms of sanguinarine on the competition between Raphidiopsis raciborskii (Cyanophyta) and Scenedesmus obliquus (Chlorophyta): A comparative toxicological study.

The phytogenic algicide sanguinarine shows strong inhibitory effects on some bloom-forming cyanobacteria and exhibits great potential in cyanobacterial bloom mitigation. To evaluate the possible ecological effects of sanguinarine on microalgae, the effects and possible mechanisms of sanguinarine on the competition between bloom-forming cyanobacterium Raphidiopsis raciborskii (formerly named Cylindrospermopsis raciborskii) and green alga Scenedesmus obliquus were investigated through co-culture competition test and comparative toxicological study including growth characteristics, chlorophyll fluorescence transients, activities of antioxidant enzymes, and lipid peroxidation. The results of Raphidiopsis-Scenedesmus co-culture competition test showed that sanguinarine decreased the competition ability of R. raciborskii, which benefitted S. obliquus in winning the competition. Toxicological studies have shown that sanguinarine exhibited high inhibitory effects on the growth and photosynthesis of R. raciborskii but no obvious toxicity on S. obliquus at concentrations of no more than 80 µg L-1. Oxidative damage partially contributed but was not the primary mechanism for the toxicity of sanguinarine on R. raciborskii. The results presented in this study indicate that sanguinarine may be a good algicidal candidate in mitigation of Raphidiopsis-based water bloom.

Life-history responses of Daphnia sinensis simultaneously exposed to Microcystis aeruginosa and Cylindrospermopsis raciborskii.

Many cyanobacterial species co-occur commonly in a freshwater ecosystem and can be consumed simultaneously by zooplankton. Both Microcystis aeruginosa and Cylindrospermopsis raciborskii are the dominant species and coexist in eutrophic tropical waters, and they together are assumed to have exert a stronger effect on the life history traits of cladocerans than a single cyanobacterial species. In the present study, we tested the hypothesis with life-history experiments of Daphnia sienesis, a large cladoceran species in tropics. In the experiments, M. aeruginosa and C. raciborskii were used as a mixture of food with Chlorella pyrenoidosa for the experimental animals. D. sienesis showed excellent growth and survival on sole diets of C. pyrenoidosa (CP). By contrast, Daphnia's growth decreased significantly and reproduction was completely inhibited when cyanobacteria comprised 100% of the food offered. The supplementation of C. pyrenoidosa into cyanobacterial diets significantly decreased their harmful effects on Daphnia, who improved the life history traits with the reduction of cyanobacterial percentage in mixed foods, irrespective of cyanobacterial species. Compared with the cultures of D. sienesis fed with a single cyanobacteria species, the animals in the treatments fed both M.aeruginosa-FACHB469 (F469) and C. raciborskii N8 (N8) had a lower growth rate in all the proportion of C. pyrenoidosa (25% CP, 50% CP or 75% CP). The strongest synergistic inhibition by the two cyanobacterial strains was found in the treatments with the 25% CP (i.e., 25% CP + 37.5% F469 + 37.5% N8), and no animal survived to maturity and reproduced. Thus, the simultaneous exposure to the two cyanobacterial species should be taken into account in assessing the ecological risks of cyanobacterial blooms, since multiple cyanobacterial coexistence can result in strong synergistic inhibition on growth and reproduction of zooplankton.

Species diversity and seasonal dynamics of filamentous cyanobacteria in urban reservoirs for drinking water supply in tropical China.

Filamentous cyanobacteria have been observed to become the dominant species in reservoirs, especially in small reservoirs for drinking water supply in southern China. The occurrences of filamentous cyanobacteria blooms in such reservoirs add additional costs for water plants by decreasing the filtration efficiency and the potential of toxin production. To serve the purpose of drinking water supply, the effective risk assessment requires the dynamic pattern of filamentous cyanobacteria. This study seasonally collected samples from 25 reservoirs in Dongguan, one of the most important 'world factories' in China in July, December and March, and investigated the temporal dynamics of phytoplankton, particularly cyanobacteria community. Our investigation showed that filamentous cyanobacteria, Planktothrix sp, Limnothrix sp. and Cylindrospermopsis raciborskii dominated in these reservoirs and climate-related water temperature was the primary factor for the seasonal shift of filamentous cyanobacteria. High abundance of filamentous cyanobacteria occurred in the high water level period with increasing temperature but less relevant with nutrient conditions. Our study observed the seasonal dynamics of filamentous cyanobacteria in tropical urban reservoirs and highlighted the association between temperature and filamentous cyanobacteria. our data and analysis provided an evidence that increased temperature could increase the likelihood of frequency and intensity of filamentous cyanobacteria blooms. In the scenario of global warming, more frequent monitoring of filamentous cyanobacteria and the potential to produce toxin should be considered for water quality and reservoir management.

Alternative Isolation Protocol for Desulfo and Zwitterionic Cylindrospermopsin Alkaloids and Comparison of Their Toxicity in HepG2 Cells.

The term cylindrospermopsins (CYNs) refers to a structurally related class of cyanobacterial metabolites comprised of a tricyclic guanidine group and a hydroxymethyluracil moiety. Most reports in environmental aquatic samples refer to cylindrospermopsin (CYN), and reports on other CYN alkaloids are scarce, due, in part, to a lack of versatile isolation protocols. Thus, using commercially available solid phase extraction (SPE) cartridges, we optimized an isolation protocol for the complete recovery of CYN, 7-deoxy-cylindrospermopsin (7D-CYN) and 7-deoxy-desulfo-cylindrospermopsin (7D-desulfo-CYN) from the same aliquot. The isolation protocol was adaptable depending on the nature of the sample (solid biomass, culture broth or environmental water sample) and tolerates up to 4 L of dense culture broth or 400 mg of lyophilized biomass. To quantitate the CYN alkaloids, we validated an LC-DAD-MS2 method, which takes advantage of the UV absorption of the uracil group (λ 262 nm). Using electrospray ionization (ESI) in a positive ion mode, the high-resolution MS1 data confirms the presence of the protonated alkaloids, and the MS2 fragment assignment is reported as complementary proof of the molecular structure of the CYNs. We isolated three CYN alkaloids with different water solubility using the same lyophilized sample, with a purity that ranged from 95% to 99%. The biological activity of the purified CYNs, along with a synthetic degradation product of CYN (desulfo-cylindrospermopsin), was evaluated by assessing necrosis and apoptosis in vitro using flow cytometry. CYN's lethal potency in HepG2 cells was greater than the other analogs, due to the presence of all four functional groups: guanidine, uracil, C-7 hydroxyl and the sulfate residue.

Physiological responses of the freshwater N2 -fixing cyanobacterium Raphidiopsis raciborskii to Fe and N availabilities.

The cyanobacterium Raphidiopsis raciborskii is of environmental and social concern in view of its toxicity, bloom-forming characteristics and increasingly widespread occurrence. However, while availability of macronutrients and micronutrients such as N and Fe are critically important for the growth and metabolism of this organism, the physiological response of toxic and non-toxic strains of R. raciborskii to varying Fe and N availabilities remains unclear. By determining physiological parameters as a function of Fe and N availability, we demonstrate that R. raciborskii growth and N2 -fixing activity are facilitated at higher Fe availability under N2 -limited conditions with faster growth of the CS-506 (cylindrospermopsin-producing) strain compared with that of CS-509 (the non-toxic) strain. Radiolabelled Fe uptake assays indicated that R. raciborskii acclimated under Fe-limited conditions acquires Fe at significantly higher rates than under Fe replete conditions, principally via unchelated Fe(II) generated as a result of photoreduction of complexed Fe(III). While N2 -fixation of both strains occurred during both day and night, the CS-506 strain overall exhibited higher N2 -fixing and Fe uptake rates than the CS-509 strain under N-deficient and Fe-limited conditions. The findings of this study highlight that Fe availability is of significance for the ecological advantage of CS-506 over CS-509 in N-deficient freshwaters.

Chemistry of a Unique Polyketide-like Synthase.

Like many complex natural products, the intricate architecture of saxitoxin (STX) has hindered full exploration of this scaffold's utility as a tool for studying voltage-gated sodium ion channels and as a pharmaceutical agent. Established chemical strategies can provide access to the natural product; however, a chemoenzymatic route to saxitoxin that could provide expedited access to related compounds has not been devised. The first step toward realizing a chemoenzymatic approach toward this class of molecules is the elucidation of the saxitoxin biosynthetic pathway. To date, a biochemical link between STX and its putative biosynthetic enzymes has not been demonstrated. Herein, we report the first biochemical characterization of any enzyme involved in STX biosynthesis. Specifically, the chemical functions of a polyketide-like synthase, SxtA, from the cyanobacteria Cylindrospermopsis raciborskii T3 are elucidated. This unique megasynthase is comprised of four domains: methyltransferase (MT), GCN5-related N-acetyltransferase (GNAT), acyl carrier protein (ACP), and the first example of an 8-amino-7-oxononanoate synthase (AONS) associated with a multidomain synthase. We have established that this single polypeptide carries out the formation of two carbon-carbon bonds, two decarboxylation events and a stereospecific protonation to afford the linear biosynthetic precursor to STX (4). The synthetic utility of the SxtA AONS is demonstrated by the synthesis of a suite of α-amino ketones from the corresponding α-amino acid in a single step.

Carbon acquisition characteristics of six microalgal species isolated from a subtropical reservoir: potential implications for species succession.

CO2 levels in freshwater systems can fluctuate widely, potentially influencing photosynthetic rates and growth of phytoplankton. Given the right conditions, this can lead to bloom formation and affect water quality. This study investigated the acquisition of dissolved inorganic carbon (DIC) by six species of microalgae, a cyanobacterium Cylindrospermopsis raciborskii, the diatoms Cyclotella sp., Nitzschia sp., and the green algae Stichococcus sp., Staurastrum sp., and Monoraphidium sp., all isolated from a subtropical reservoir in Australia. Carbon acquisition characteristics, specifically the affinity for DIC, internal pH, and internal DIC concentrations were measured. Affinities for CO2 ( K0.5(CO2) ) ranged between 0.7 and 6 µM CO2 . This was considerably lower than air-equilibrated surface water CO2 concentrations, and below reported affinities for CO2 of RuBisCO suggesting operation of active carbon dioxide concentrating mechanisms (CCMs) in all species. Internal pH was lowest for Cyclotella sp. at 7.19, and highest for Staurastrum sp., at 7.71. At 180 µM external DIC, ratios of internal:external CO2 ranged from 2.5 for Nitzschia sp. to 14 in C. raciborskii. Internal HCO3- concentration showed a linear relationship with surface area to biovolume ratio (SA:Vol). We hypothesized that species with a higher SA:Vol suffer more from diffusive escape of CO2 , thus storage of DIC as bicarbonate is favored in these strains. For C. raciborskii, under stratified summer conditions, its strong CCM, and resilient photosynthetic characteristics may contribute to its bloom forming capacity.

The Effect of Bisphenol A on Growth, Morphology, Lipid Peroxidation, Antioxidant Enzyme Activity, and PS II in Cylindrospermopsis raciborskii and Scenedesmus quadricauda.

To investigate the effect of bisphenol A (BPA) on Cylindrospermopsis raciborskii (Cyanobacteria) and Scenedesmus quadricauda (Chlorophyta), we grew the two species at BPA concentrations of 0, 0.1, 1, 2, 5, 10, and 20 mg/L and examined their growth, lipid peroxidation, antioxidant enzyme activity, and chlorophyll a fluorescence. The 96-h EC50 values (effective concentration causing 50% growth inhibition) for BPA in C. raciborskii and S. quadricauda were 9.663 ± 0.047, and 13.233 ± 0.069 mg/L, respectively. A significant reduction in chlorophyll a concentration was found in C. raciborskii and S. quadricauda when BPA concentrations were greater than 1 and 2 mg/L, respectively. Furthermore, F v/F m, ΔF/F m', and qP decreased significantly at 10 mg/L BPA in C. raciborskii but started to decrease at 10 mg/L in S. quadricauda. The changes in chlorophyll fluorescence parameters (α, rETRmax) that were obtained from the rapid light response curves of both algae species showed similar responses to F v/F m, ΔF/F m', and qP under BPA-induced stress. Values for all of the chlorophyll fluorescence parameters in S. quadricauda were higher than in C. raciborskii; however, the nonphotochemical quenching measured in C. raciborskii was considerably higher than it was in S. quadricauda. In addition, lipid peroxidation (determined as MDA content) and antioxidant enzyme activities (SOD and CAT) increased in both species as the BPA concentration increased. These results suggest that C. raciborskii is more sensitive to the effects of BPA than S. quadricauda and that photosystem II might be a target for the activity of BPA in vivo.

First report of neurotoxic effect of the cyanobacterium Cylindrospermopsis raciborskii on the motility of trematode metacercariae.

Cylindrospermopsis raciborskii (Woloszynska) is a photosynthetic cyanobacterium that can produce cytotoxic (cylindrospermopsin) and neurotoxic cyanotoxins (saxitoxins). In Brazil the strains of C. raciborskii are reported to produce only saxitoxins (STX) and their effect on fish parasites has not been tested to date. The fish Poecilia vivipara Bloch and Schneider is a common host for the trematode Pygidiopsis macrostomum Travassos off the coast of Rio de Janeiro, and this fish-parasite interaction is a model for behavioural and ecotoxicological studies. The aim of this work was to evaluate the motility of metacercariae of P. macrostomum from P. vivipara exposed to 40 mg l-1 and 400 mg l-1 of crude lyophilized extract of the cyanobacterium C. raciborskii (CYRF-01) for 48 h. The fish were separated into groups of ten individuals and, after exposure, five fish from each group were dissected for counting and checking the motility of metacercariae. The other five fish were dissected after 48 h in clean water. The detection and quantification of STX in the solutions of cyanobacteria, and the gills and guts of fish, were performed by an enzyme-linked immunosorbent assay. The crude extract of C. raciborskii caused temporary paralysis in metacercariae of P. macrostomum after exposure of fish to both concentrations, and the motility recovered after the fish were kept for 48 h in clean water. STX was detected in the guts and gills of all fish analysed, suggesting that this toxin is involved in the paralysis of metacercariae. This is the first report on the action of neurotoxins in metacercariae of fish.

The effects of interspecific interactions between bloom forming cyanobacteria and Scenedesmus quadricauda (chlorophyta) on their photophysiology.

Eutrophication and enhanced external nutrient loading of lakes and seas are most clearly reflected by increased cyanobacterial blooms, which are often toxic. Freshwater cyanobacteria produce a number of bioactive secondary metabolites, some of which have allelopathic properties, significantly influencing the biological processes of other algae, thereby affecting species composition and succession of the phytoplankton. The goal of this work was to investigate the influence of bloom-forming cyanobacterial exudates on the photophysiology of the green alga Scenedesmus quadricauda by chlorophyll fluorescence analysis. We were able to prove the effect of algal cell-free filtrates on the performance of S. quadricauda and demonstrate for the first time that the freshwater picocyanobacterium Cyanobium gracile has strong negative impact on the coexisting green alga. Neither the cyanotoxin (MYC, CYN and ATX) producing, nor the non-toxic strains showed any systematic effect on the production of S. quadricauda. Various strains of the cyanobacterium Cylindrospermopsis raciborskii inhibited the performance of the green alga independently of their origin. Our results urge further studies for a better understanding of the factors affecting the release of allelopathic compounds and the mechanisms of their effects on target organisms.

Influence of environmental variables on saxitoxin yields by Cylindrospermopsis raciborskii in a mesotrophic subtropical reservoir.

Saxitoxins are a class of toxins produced by at least two groups of evolutionarily distant organisms (cyanobacteria and dinoflagellates). While the toxicity of these toxins is relatively well characterized, to date little is known about their drivers and ecological functions, especially in lower latitude tropical and subtropical freshwater ecosystems. In the present study, we aimed to obtain a better understanding of the main drivers of saxitoxin concentrations in aquatic environments. We investigated the relationships among saxitoxin concentrations in a mesotrophic subtropical reservoir dominated by the cyanobacteria Cylindrospermopsis raciborskii with physical, chemical and biological water variables. The highest saxitoxin concentrations were 0.20 µg·L-1, which occurred in the samples with the highest densities of C. raciborskii (maximum of 4.3 × 104 org·mL-1) and the highest concentration of dissolved nutrients (nitrate from 0.2 to 0.8 µg·L-1, ortophosphate from 0.3 to 8.5 µg·L-1). These correlations were confirmed by statistical analyses. However, the highest saxitoxin relative concentrations (per trichome) were associated with lower C. raciborskii densities, suggesting that saxitoxin production or the selection of saxitoxin-producing strains was associated with the adaptation of this species to conditions of stress. Our results indicate that C. raciborskii toxin yields vary depending on the enrichment conditions having potential implications for reservoir management.

The occurrence of toxic cyanobacterium Cylindrospermopsis raciborskii and its toxin cylindrospermopsin in the Huong River, Thua Thien Hue province, Vietnam.

This research reports the presence of species Cylindrospermopsis raciborskii and cylindrospermopsin (CYN) in the Huong River and the relationship between species with environmental factors to find a scientific basis for predicting the risk of pollution of the species and CYN in waters. Strains of C. raciborskii isolated from the river were also identified as potentially toxin-producing through the determination of the presence of toxins in the cultures by ELISA; the presence of the genes involved by PCR confirms the CYN-producing ability of species C. raciborskii from this water body. Our results have confirmed the presence of toxic cyanobacteria C. raciborskii in the Huong River. C. raciborskii from the Huong River are mostly solitary, straight trichomes. Analyses of all C. raciborskii strains from the Huong River by ELISA for cylindrospermopsin were positive. The contents of cylindrospermopsin (CYN) in each strain were different, ranging from 5.25 ng mg-1 wet weight in CR1DD to 70.83 ng mg-1 wet weight in CR1NY. PCR analysis confirmed that the genes involved in the production of this cyanotoxin were present in C. raciborskii. The relationship between densities and toxicity showed a correlation coefficient R of 0.88. This was a relatively high positive correlation index, indicating the close links between densities and toxins: toxin CYN concentrations increased when C. raciborskii densities increased.