Presence Of Cylindrospermopsin Research Articles

Snapshot of cyanobacterial toxins in Pakistani freshwater bodies.

Cyanobacteria are known to produce diverse secondary metabolites that are toxic to aquatic ecosystems and human health. However, data about the cyanotoxins occurrence and cyanobacterial diversity in Pakistan's drinking water reservoirs is scarce. In this study, we first investigated the presence of microcystin, saxitoxin, and anatoxin in 12 water bodies using an enzyme-linked immunosorbent assay (ELISA). The observed cyanotoxin values for the risk quotient (RQ) determined by ELISA indicated a potential risk for aquatic life and human health. Based on this result, we made a more in-depth investigation with a subset of water bodies (served as major public water sources) to analyze the cyanotoxins dynamics and identify potential producers. We therefore quantified the distribution of 17 cyanotoxins, including 12 microcystin congeners using a high-performance liquid chromatography-high-resolution tandem mass spectrometry/mass spectrometry (HPLC-HRMS/MS). Our results revealed for the first time the co-occurrence of multiple cyanotoxins and the presence of cylindrospermopsin in an artificial reservoir (Rawal Lake) and a semi-saline lake (Kallar Kahar). We also quantified several microcystin congeners in a river (Panjnad) with MC-LR and MC-RR being the most prevalent and abundant. To identify potential cyanotoxin producers, the composition of the cyanobacterial community was characterized by shotgun metagenomics sequencing. Despite the noticeable presence of cyanotoxins, Cyanobacteria were not abundant. Synechococcus was the most abundant cyanobacterial genus found followed by a small amount of Anabaena, Cyanobium, Microcystis, and Dolichospermum. Moreover, when we looked at the cyanotoxins genes coverage, we never found a complete microcystin mcy operon. To our knowledge, this is the first snapshot sampling of water bodies in Pakistan. Our results would not only help to understand the geographical spread of cyanotoxin in Pakistan but would also help to improve cyanotoxin risk assessment strategies by screening a variety of cyanobacterial toxins and confirming that cyanotoxin quantification is not necessarily related to producer abundance.

Evaluation of the Removal and Effects of Cylindrospermopsin on Ripened Slow Sand Filters.

The occurrence of toxic blooms of cyanobacteria has been a matter of public health interest due to the cyanotoxins produced by these microorganisms. Cylindrospermopsin (CYN) is a cyanotoxin of particular concern due to its toxic effects on humans. This study investigated the removal and effects of CYN in ripened slow sand filters (SSFs) treating water from Paranoá Lake, Brasilia, Brazil. Four pilot-scale SSFs were ripened and operated for 74 days. Two contamination peaks with CYN were applied along the filtration run. The improvement of any of the evaluated water quality parameters was not affected by the presence of CYN in the raw water. The SSFs efficiently removed CYN, presenting concentrations lower than 0.8 µg/L in the filtered water. The microbiota of the SSFs were dominated by protozoa of the genus Euglypha and amoebas of the genera Arcella, Centropyxis, and Amoeba, together with some groups of rotifers. These microorganisms played a crucial role in removing total coliforms and E. coli. In addition, CYN was not identified as a determining factor in the microbiota composition.

Cytotoxic Effects and Oxidative Stress Produced by a Cyanobacterial Cylindrospermopsin Producer Extract versus a Cylindrospermopsin Non-Producing Extract on the Neuroblastoma SH-SY5Y Cell Line.

The incidence and interest of cyanobacteria are increasing nowadays because they are able to produce some toxic secondary metabolites known as cyanotoxins. Among them, the presence of cylindrospermopsin (CYN) is especially relevant, as it seems to cause damage at different levels in the organisms: the nervous system being the one most recently reported. Usually, the effects of the cyanotoxins are studied, but not those exerted by cyanobacterial biomass. The aim of the present study was to assess the cytotoxicity and oxidative stress generation of one cyanobacterial extract of R. raciborskii non-containing CYN (CYN-), and compare its effects with those exerted by a cyanobacterial extract of C. ovalisporum containing CYN (CYN+) in the human neuroblastoma SH-SY5Y cell line. Moreover, the analytical characterization of potential cyanotoxins and their metabolites that are present in both extracts of these cultures was also carried out using Ultrahigh Performance Liquid Chromatography-Mass Spectrometry, in tandem (UHPLC-MS/MS). The results show a reduction of cell viability concentration- and time-dependently after 24 and 48 h of exposure with CYN+ being five times more toxic than CYN-. Furthermore, the reactive oxygen species (ROS) increased with time (0-24 h) and CYN concentration (0-1.11 µg/mL). However, this rise was only obtained after the highest concentrations and times of exposure to CYN-, while this extract also caused a decrease in reduced glutathione (GSH) levels, which might be an indication of the compensation of the oxidative stress response. This study is the first one performed in vitro comparing the effects of CYN+ and CYN-, which highlights the importance of studying toxic features in their natural scenario.

Ferrate(VI) mediated degradation of the potent cyanotoxin, cylindrospermopsin: Kinetics, products, and toxicity

The presence of cylindrospermopsin (CYN), a potent cyanotoxin, in drinking water sources poses a tremendous risk to humans and the environment. Detailed kinetic studies herein demonstrate ferrate(VI) (FeVIO42−, Fe(VI)) mediated oxidation of CYN and the model compound 6-hydroxymethyl uracil (6-HOMU) lead to their effective degradation under neutral and alkaline solution pH. A transformation product analysis indicated oxidation of the uracil ring, which has functionality critical to the toxicity of CYN. The oxidative cleavage of the C5=C6 double bond resulted in fragmentation of the uracil ring. Amide hydrolysis is a contributing pathway leading to the fragmentation of the uracil ring. Under extended treatment, hydrolysis, and extensive oxidation lead to complete destruction of the uracil ring skeleton, resulting in the generation of a variety of products including nontoxic cylindrospermopsic acid. The ELISA biological activity of the CYN product mixtures produced during Fe(VI) treatment parallels the concentration of CYN. These results suggest the products do not possess ELISA biological activity at the concentrations produced during treatment. The Fe(VI) mediated degradation was also effective in the presence of humic acid and unaffected by the presence of common inorganic ions under our experimental conditions. The Fe(VI) remediation of CYN and uracil based toxins appears a promising drinking water treatment process.

Effect of cold food storage techniques on the contents of Microcystins and Cylindrospermopsin in leaves of spinach (Spinacia oleracea) and lettuce (Lactuca sativa).

The presence of Cylindrospermopsin (CYN) and Microcystins (MCs) in vegetables is considered as a significant worldwide toxicological risk. Thus, this work aims to assess for the first time the impact of refrigeration (4°C) and freezing (-20°C) on the levels of CYN, MCs and their mixtures (CYN+MCs) in lettuce and spinach. Samples were spiked with 750μg cyanotoxins/g dry weight (d.w.). Several storage conditions were studied: refrigeration after 24, 48h and 7 days, and freezing for 7 days, 1 and 3 months. Cyanotoxin concentrations were determined by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS). For CYN, refrigeration at 48h and 7 days was effective to decrease its concentrations up to 26% and 32%, respectively, in spinach. For MCs, refrigeration was only effective in lettuce compared to spinach, showing an important decrease of 80.3% MC-LR and 85.1% MC-YR. In spinach, CYN was stable after 3 months freezing, whereas MC contents were still reduced up to 44%. Overall, cyanotoxins were less stable in the mixture compared to individual toxins for both processes, and the effect of these storage techniques were toxin and food-specific. Further studies of cyanotoxins in foods are required for evaluating the risk for humans.

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.

First occurrence of Cylindrospermopsin in the Azores (Lake São Brás, S. Miguel Island)

Anthropogenic impacts have been reported as one of the main causes of eutrophication in freshwater systems and nutrient availability, mainly from discharges, are key to cyanobacteria development and harmful cyanobacteria blooms (cyanoHABs) upsurge. As these scenarios are common in the Azores, a cyanobacteria monitoring program was implemented in the lakes of the archipelago, which comprises cyanobacteria identification and cyanotoxin (only microcystin) analysis in over 15 lakes. However other cyanotoxins should be searched as is the case of cylindrospermopsin (CYN). This cyanotoxin has been reported more recently in a wider range of countries, because of global warming. Using ESI–LC–MS/MS (electrospray ionization–liquid chromatography–mass spectrometry), the presence of CYN was investigated in samples from four eutrophic lakes. Cylindrospermopsin was detected in one sample from Lake Sao Bras, which represents a threat to environmental and public health. This is the first CYN identification in the Azores reinforcing the necessity for lake monitoring. We propose the use of a polyphasic approach to fully understand CYN presence in the Azorean lakes, investigating the producing taxa, the environmental factors that promote cyanobacteria growth, and CYN toxicity effects.

Probing high-affinity aptamer binding region and development of aptasensor platform for the detection of cylindrospermopsin

Cylindrospermopsin (CYN) is one of the most concerning cyanotoxins due to its potential toxicity and spreading to various environments including drinking water. CYN has potential interferences with human and animal metabolic pathways, which influence the functions of organs including liver, kidneys, lungs, etc. CYN is involved in the inhibition of protein synthesis and detachment of ribosomes from the endoplasmic reticulum membrane. It also interacts with soluble proteins, which are associated with protein translations. It is believed that cytochrome 450 is responsible for the rapid toxicity of CYN. Researchers are urged to develop a high-throughput screening method for the detection of CYN in water. Construction of low cost, rapid, and sensitive analytical methods for the detection of CYN is challenging. Here, we used graphene oxide (GO) as the fluorescence sensing platform for probing the high affinity of the short aptamer derived from the wild-type long aptamer-CYN sensing. The biosensor construction involved two steps: first, quenching the fluorescence of fluorescent-labelled truncated aptamer using GO as a quencher and, second, fluorescence recovery in the presence of CYN by competitive binding between the target and GO. One of the truncate aptamers has a 12-fold higher affinity and enhances sensitivity compared to the long aptamer sequence. The limit of detection of the high affinity truncated aptamer is 17pM which is 6-fold lower than the long aptamer (100pM). The sensor specifically detects CYN in the presence of other potential interfering toxins. The performance of the sensor was validated using CYN spiked tap water with very good recovery percentage. A rapid and highly sensitive detection of CYN from water resources has been achieved using this method.

Quantification of cylindrospermopsin, anatoxin-a and homoanatoxin-a in cyanobacterial bloom freshwater using direct injection/SPE coupled with UPLC-MS/MS

Analytical methods based on direct injection (DI) and solid phase extraction (SPE) coupled with ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC- MS/MS) were developed for the determination of anatoxin-a (ATX-a), cylindrospermopsin (CYN), and homoanatoxin-a (HATX-a) in freshwater samples impacted with cyanobacterial blooms. The presence of CYN in freshwater samples was detected and quantified based on direct injection method, while ATX-a and HATX-a could be determined by both DI and SPE-based methods. Matrix effects (ME) on the signal intensity of the cyanotoxins were systematically evaluated for both direct injection and SPE extract samples. CYN, ATX-a, and HATX-a suffered a significant suppression during UPLC-MS/MS. The selection of internal standards (ISs) for compensating/correcting the losses of target cyanotoxins during sample preparation and matrix effects in UPLC-MS/MS analyses were systematically evaluated. Acetaminophen-d4 (an isotopically labelled acetaminophen) is a suitable internal standard for correcting the ME on the signal intensity of ATX-a and HATX-a, while the use of L-phenylalanine-d5 or caffeine-d9 as IS for correcting ME of these toxins was not efficient, as expected. The method detection limit (MDL) for the target cyanotoxins ranged from 0.6 to 15 ng/L, which is sensitive enough to detect the presence of these toxins in cyanobacterial bloom freshwater. The developed methods were successfully applied for routine monitoring of the occurrence of these cyanotoxins in a local water body. Monitoring results depicted that ATX-a, CYN and HATX-a were ubiquitously detected in water samples, at concentrations ranging from 70 to 24,600 ng/L.

Impact of the toxicity of Cylindrospermopsis raciborskii (Woloszynska) Seenayya & Subba Raju on laboratory rats in vivo.

In vivo laboratory studies of toxicity were performed on Wistar rats using a methanol extract produced by the natural population of Cylindrospermopsis raciborskii (abundance of 2.13×105 trichomesmL-1) collected at Aleksandrovac Lake (Serbia). HPLC analysis showed that the extract contains 6.65μg cylindrospermopsin (CYN) mg-1. The rats were killed 24 or 72h after a single intraperitoneal injection of C. raciborskii extract in concentrations of 1500, 3000, 6000 and 12,000μgkg-1 body weight (bw) and an equivalent amount of CYN as present in the highest dose of the extract (79.80μg CYNkg-1 bw). The genotoxic effect on the livers treated with C. raciborskii was evaluated using comet assay and potential induction of oxidative stress as the toxicity mechanism associated with the presence of CYN in extract. The results from the analyses of DNA damage in the comet tail length, tail moment and percentage of DNA in the tail in the liver indicated that administration of extract and CYN present statistically significant difference when compared with the negative control group. Although an increase in the frequency of selected parameters induced by the CYN was observed in the liver, this damage was less than the damage resulting from the administration of the highest dose of extract. The changes in the biochemical parameters of the hepatic damage showed that the application of single doses of the extract and CYN did not cause serious liver damage in rats. The extract and CYN significantly increased oxidative stress in rats' liver after a single exposure.

Changes on cylindrospermopsin concentration and characterization of decomposition products in fish muscle (Oreochromis niloticus) by boiling and steaming

The occurrence of cylindrospermopsin (CYN) in toxic blooms of cyanobacteria is increasing in both frequency and distribution over the world. CYN persistence in water potentiates its accumulation in a wide range of aquatic animals destined to human consumption representing a serious problem. The aim of this work was to study for the first time the influence of cooking (boiling and steaming) for 1 or 2 min, on the stability of CYN in muscle of fish (Oreochromis niloticus) contaminated under laboratory conditions, also analyzing the waters in which samples were cooked. CYN was extracted and quantification was carried out by a developed and validated UPLC-MS/MS method. The results show that concentrations of CYN in fish are dependent on the cooking method, being the steaming for 2 min the most effective in reducing CYN levels (26%), followed by boiling for 2 min (18%), and significant differences have been found between the two periods assayed (1 min vs. 2 min). CYN was also detected in waters in which fish muscles were cooked in the range 0.10–0.28 μg/L. Moreover, characteristic decomposition products depending on the type of cooking were detected for the first time among the results of these treatments. The present findings emphasize the need for further studies to evaluate the influence of cooking in the presence of CYN in fish for a more realistic risk evaluation for the human health.

Cylindrospermopsin Biodegradation Abilities of Aeromonas sp. Isolated from Rusałka Lake.

The occurrence of the cyanobacterial toxin cylindrospermopsin (CYN) in freshwater reservoirs is a common phenomenon. However, the biodegradation of this toxin in environmental samples has been observed only occasionally. In this work the biodegradation ability of cylindrospermopsin was investigated based on isolates from lakes with previous cyanotoxin history. Bacterial strains were identified based on the 16S rDNA and rpoD gene comparison. CYN biodegradation was monitored using the HPLC method. The R6 strain identified as Aeromonas sp. was documented as being capable of CYN removal. This biodegradation was dependent on the pH and temperature. Additionally, the stimulation of the growth of the R6 strain in the presence of CYN was indicated. Our discovery supports the hypothesis that (in analogy to the well-known phenomenon of microcystin biodegradation) in lakes dominated by potential CYN-producing cyanobacteria, the processes of microbial utilization of this toxin may occur.

Biochemical and growth performance of the aquatic macrophyte Azolla filiculoides to sub-chronic exposure to cylindrospermopsin.

Physiological and biochemical effects of cylindrospermopsin (CYN), a cyanobacterial toxin that inhibits protein synthesis and released during a harmful cyanobacterial bloom, has been overlooked in plants. Therefore, at the present research, the toxic effects (physiological and biochemical) of a crude extract containing CYN were assessed in the aquatic fern Azolla filiculoides exposed to three concentrations (0.05, 0.5 and 5 μg CYN mL(-1)). At 5 μg CYN mL(-1), fern growth rate has showed a drastic decrease (0.001 g g(-1) day(-1)) corresponding to a 99.8% inhibition, but at the concentrations of 0.05 and 0.5 μg CYN mL(-1) the growth rate was similar to the control plants. Growth rate also indicated a IC50 of 2.9 μg CYN mL(-1). Those data point to the presence of other compounds in the crude extract may stimulate the fern growth and/or the fern is tolerant to CYN. Chlorophyll (a and b), carotenoids and protein content as well as the activities of glutathione reductase (GR) and glutathione-S-transferase (GST) has increased at 5 μg CYN mL(-1) which may indicate that photosynthesis and protein synthesis are not affected by CYN and the probable activation of defense and detoxifying mechanisms to overcome the effects induced by the presence of CYN. Low uptake of cylindrospermopsin (1.314 μg CYN g(-1) FW) and low bioconcentration factor (0.401) point towards to a safe use of A. filiculoides as biofertilizer and as food source, but also indicate that the fern is not suitable for CYN phytoremediation.

Biodegradation of cylindrospermopsin toxin by microcystin-degrading bacteria isolated from cyanobacterial blooms

Cylindrospermopsin (CYN) is highly water soluble and stable cyanotoxin which can persist in water sources and pose health hazards to human, animals and plants. Consequently, there is a need to fully understand the fate of CYN in the aquatic environment, in particular, its biodegradation by endemic organisms. Although there is evidence of CYN degradation by microbial population, bacterial strains responsible for the toxin degradation are not yet identified. This study reports for the first time CYN degradation by Bacillus strain (AMRI-03) isolated from cyanobacterial blooms. In degradation batch experiment, the strain grew well in the presence of CYN without showing any lag period, and this growth increased with the initial CYN concentration. CYN degradation occurred rapidly, and the complete degradation was dependent on the initial CYN concentration. It occurred after 6 days at the highest concentration (300 μg L−1) compared to 7 and 8 days at lower concentrations (10 & 100 μg L−1). Also, the degradation rate correlated positively with the initial CYN concentration with maximum value (50 μg L−1 day−1) obtained at the highest CYN concentration. Furthermore, the biodegradation rate of CYN by this strain depended remarkably on temperature and pH. The highest biodegradation rates were obtained at 25 and 30 °C, and at pH 7 and pH 8. Taken that such microcystin-degrading strain can also degrade CYN, bacterial strains reported worldwide as microcystin degraders could be tested for their capability of CYN degradation.

Preliminary molecular identification of cylindrospermopsin-producing Cyanobacteria in two Polish lakes (Central Europe)

The presence of toxigenic cyanobacteria capable of biosynthesis of cylindrospermopsin (CYN) was measured in 24 water samples collected from the lakes Bytyńskie (BY) and Bnińskie (BN) in the Western Poland. The study also covered analysis of toxigenicity and production of CYN by the culture of Cylindrospermopsis raciborskii isolated from BY. The cyrJ gene associated with CYN production was identified in 22 water samples collected in the summer seasons of 2006 and 2007. The presence of CYN was confirmed in 16 samples. The homology searches revealed that amplified sequences of four water samples, which were selected from among all the samples, displayed a strong 99% homology to cyrJ gene of Aphanizomenon sp. 10E6. The culture of C.raciborskii did not contain the cyrJ gene nor the CYN. The specificity of C.raciborskii was confirmed by application of a fragment of the rpoC1. These first genetic analyses have shown that Aphanizomenon seems to be the main cyanobacterial genus responsible for the production of CYN in the Polish lakes. The lack of toxigenicity of the isolated C.raciborskii suggests that it is possible that this invasive species does not demonstrate toxigenic activity in Polish water bodies.

Novel toxic effects associated with a tropical Limnothrix/Geitlerinema‐like cyanobacterium

The presence of a toxic strain of a fine filamentous cyanobacterium belonging to the Oscillatorialean family Pseudanabaenacea was detected during a survey of cyanobacterial taxa associated with the presence of cylindrospermopsin in dams in Central Queensland (Australia). The strain, AC0243, was isolated and cultured, its genomic DNA extracted and 16S RNA gene sequenced. Phylogenetic analysis placed AC0243 with Limnothrix species, although this genus appears polyphyletic. Moreover, not all morphological characters are consistent with this genus but more closely fit the description of Geitlerinema unigranulatum (R.N. Singh) Komárek and Azevedo. The potential toxic effects of AC0243 extract were assessed chemically and biologically. Cell free protein synthesis was inhibited by the extract. Exposure of Vero cells to the extract resulted in a significant reduction in cellular ATP levels following 24-72 h incubation. The presence of cylindrospermopsin was excluded based on the nature of responses obtained in cell and cell-free assays; in addition, (i) it could not be detected by HPLC, LC-MS, or immunological assay, and (ii) no genes currently associated with the production of cylindrospermopsin were found in the genome. Other known cyanobacterial toxins were not detected. The apparent novelty of this toxin is discussed.

Differential protein expression in two bivalve species; Mytilus galloprovincialis and Corbicula fluminea; exposed to Cylindrospermopsis raciborskii cells

The cyanobacteria Cylindrospermopsis raciborskii is considered a threat to aquatic organisms due to the production of the toxin cylindrospermopsin (CYN). Despite the numerous reports evidencing the toxic effects of C. raciborskii cells and CYN in different species, not much is known regarding the toxicity mechanisms associated with this toxin and the cyanobacteria. In this work, a proteomics approach based in the two-dimensional gel electrophoresis and mass spectrometry was used to study the effects of the exposure of two bivalve species, Mytilus galloprovincialis and Corbicula fluminea, to CYN producing (CYN+) and non-producing (CYN−) C. raciborskii cells. Additionally the activities of glutathione S-transferase (GST) and glutathione peroxidase (GPx) were determined. Alterations in actin and tubulin isoforms were detected in gills of both bivalve species and digestive gland of M. galloprovincialis when exposed to CYN− and CYN+ cells. Moreover, GST and GPx activities changed in gills and digestive tract of bivalves exposed to both C. raciborskii freeze dried cells, in comparison to control animals exposed to the green alga Chlorella vulgaris. These results suggest the induction of physiological stress and tissue injury in bivalves by C. raciborskii. This condition is supported by the changes observed in GPx and GST activities which indicate alterations in the oxidative stress defense mechanisms. The results also evidence the capacity of CYN non-producing C. raciborskii to induce biochemical responses and therefore its toxicity potential to bivalves. The heat shock protein 60 (HSP60), extrapallial (EP) fluid protein and triosephosphate isomerase homologous proteins from gills of M. galloprovincialis were down-regulated specifically with the presence of CYN+ C. raciborskii cells. The presence of CYN may lead to additional toxic effects in M. galloprovincialis. This work demonstrates that proteomics is a powerful approach to characterize the biochemical effects of C. raciborskii and to investigate the physiological condition of the exposed organisms.

First evidence of “paralytic shellfish toxins” and cylindrospermopsin in a Mexican freshwater system, Lago Catemaco, and apparent bioaccumulation of the toxins in “tegogolo” snails ( Pomacea patula catemacensis)

Exposure to cyanobacterial toxins in freshwater systems, including both direct (e.g., drinking water) and indirect (e.g., bioaccumulation in food webs) routes, is emerging as a potentially significant threat to human health. We investigated cyanobacterial toxins, specifically cylindrospermopsin (CYN), the microcystins (MCYST) and the “paralytic shellfish toxins” (PST), in Lago Catemaco (Veracruz, Mexico). Lago Catemaco is a tropical lake dominated by Cylindrospermopsis, specifically identified as Cylindrospermopsis catemaco and Cylindrospermopsis philippinensis, and characterized by an abundant, endemic species of snail ( Pomacea patula catemacensis), known as “tegogolos,” that is both consumed locally and commercially important. Samples of water, including dissolved and particulate fractions, as well as extracts of tegogolos, were screened using highly specific and sensitive ELISA. ELISA identified CYN and PST at low concentrations in only one sample of seston; however, both toxins were detected at appreciable quantities in tegogolos. Calculated bioaccumulation factors (BAF) support bioaccumulation of both toxins in tegogolos. The presence of CYN in the phytoplankton was further confirmed by HPLC-UV and LC-MS, following concentration and extraction of algal cells, but the toxin could not be confirmed by these methods in tegogolos. These data represent the first published evidence for CYN and the PST in Lago Catemaco and, indeed, for any freshwater system in Mexico. Identification of the apparent bioaccumulation of these toxins in tegogolos may suggest the need to further our understanding of the transfer of cyanobacterial toxins in freshwater food webs as it relates to human health.

Toxicity of cylindrospermopsin, and other apparent metabolites from Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum, to the zebrafish ( Danio rerio) embryo

Cyanobacteria produce a diverse array of toxic or otherwise bioactive compounds that pose growing threats to human and environmental health. We utilized the zebrafish ( Danio rerio) embryo, as a model of vertebrate development, to investigate the inhibition of development pathways (i.e. developmental toxicity) by the cyanobacterial toxin, cylindrospermopsin (CYN), as well as extracts from various isolates of Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum. CYN was toxic only when injected directly into embryos, but not by direct immersion at doses up to 50 μg/ml. Despite the dose dependency of toxicity observed following injection of CYN, no consistent patterns of developmental defects were observed, suggesting that toxic effects of CYN may not target specific developmental pathways. In contrast, direct immersion of embryos in all of the extracts resulted in both increased mortality and reproducible, consistent, developmental dysfunctions. Interestingly, there was no correlation of developmental toxicity observed for these extracts with the presence of CYN or with previously reported toxicity for these strains. These results suggest that CYN is lethal to zebrafish embryos, but apparently inhibits no specific developmental pathways, whereas other apparent metabolites from C. raciborskii and A. ovalisporum seem to reproducibly inhibit development in the zebrafish model. Continued investigation of these apparent, unknown metabolites is needed.

First report of the cyanobacterial toxin cylindrospermopsin in New Zealand

Cylindrospermopsin, a hepatotoxin produced by cyanobacteria, has been unambiguously detected in cyanobacteria collected from a recreational lake in the Wellington region of New Zealand. To our knowledge this is the first report of cylindrospermopsin in New Zealand. Cylindrospermopsin and several microcystins were identified by reversed phase high-performance liquid chromatography-mass spectrometry (LC-MS) and tandem mass spectrometry (LC-MS/MS) using an atmospheric-pressure ionization source and an ionspray interface. The presence of cylindrospermopsin in New Zealand highlights the risk of direct cyanobacterial toxin exposure in recreational waters.