The first occurrence of the cyanobacterial alkaloid toxin cylindrospermopsin in the Czech Republic as determined by immunochemical and LC/MS methods

Bláhová, Lucie; Oravec, Michal; Maršálek, Blahoslav; Šejnohová, Lenka; Šimek, Zdeněk; Bláha, Luděk

doi:10.1016/j.toxicon.2009.01.014

Cited by 98 publications

(51 citation statements)

References 29 publications

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“…7-Epi-CYN has so far been detected only in Aphanizomenon ovalisporum as a minor metabolite (2). These three closely related molecules were shown previously to be toxic using in vivo assays, and they were shown to inhibit protein synthesis with similar potencies (22,32,33).Several analytical techniques have been developed to assay concentrations of CYN in cultures or in the environment: an enzyme-linked immunosorbent assay (ELISA) and assays based on liquid chromatography coupled either to a UV detector (40) or to a mass spectrometer (3,12,13).The biosynthesis of CYN was studied by using feeding experiments (5), and a biosynthetic scheme was later proposed based on the bioinformatic analysis of the gene cluster responsible for the biosynthesis of CYN, recently identified in C. raciborskii (25). Interestingly, 7-deoxy-CYN was proposed to be a precursor of CYN, with the hydroxylation step likely being catalyzed by CyrI, a proline hydroxylase homolog.…”

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confidence: 99%

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Biosynthesis of Cylindrospermopsin and 7-Epicylindrospermopsin in Oscillatoria sp. Strain PCC 6506: Identification of the cyr Gene Cluster and Toxin Analysis

Mazmouz

Chapuis-Hugon

Mann

et al. 2010

Appl Environ Microbiol

123

117

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Cylindrospermopsin is a cytotoxin produced by Cylindrospermopsis raciborskii and other cyanobacteria that has been implicated in human intoxications. We report here the complete sequence of the gene cluster responsible for the biosynthesis of this toxin in Oscillatoria sp. strain PCC 6506. This cluster of genes was found to be homologous with that of C. raciborskii but with a different gene organization. Using an enzyme-linked immunosorbent assay and an optimized liquid chromatography analytical method coupled to tandem mass spectrometry, we detected 7-epicylindrospermopsin, cylindrospermopsin, and 7-deoxycylindrospermopsin in the culture medium of axenic Oscillatoria PCC 6506 at the following relative concentrations: 68.6%, 30.2%, and 1.2%, respectively. We measured the intracellular and extracellular concentrations, per mg of dried cells of Oscillatoria PCC 6506, of 7-epicylindrospermopsin (0.18 g/mg and 0.29 g/mg, respectively) and cylindrospermopsin (0.10 g/mg and 0.11 g/mg, respectively). We showed that these two toxins accumulated in the culture medium of Oscillatoria PCC 6506 but that the ratio (2.5 ؎ 0.3) was constant with 7-epicylindrospermopsin being the major metabolite. We also determined the concentrations of these toxins in culture media of other Oscillatoria strains, PCC 6407, PCC 6602, PCC 7926, and PCC 10702, and found that, except for PCC 6602, they all produced 7-epicylindrospermopsin and cylindrospermopsin, with the former being the major toxin, except for PCC 7926, which produced very little 7-epicylindrospermopsin. All the cylindrospermopsin producers studied gave a PCR product using specific primers for the amplification of the cyrJ gene from genomic DNA.Cyanobacteria are known to produce a wide range of secondary metabolites (19), some of which are toxic to humans and animals, and cyanobacterial blooms are frequently associated with human-or animal-poisoning events. Several classes of cyanobacterial toxins have so far been identified, including hepatotoxins, neurotoxins, cytotoxins, and irritants (42). Cases of animal intoxication due to cyanobacterial toxin exposure are regularly reported in different places around the world, and it is now recognized that the release of cyanobacterial toxins in water reservoirs and water supplies has major implications for public health and for the environment (8). For instance, anatoxin-a and homoanatoxin-a, two cyanobacterial neurotoxins, provoke the rapid death of animals by acute asphyxia when ingested, because these alkaloids are potent agonists of the nicotinic acetylcholine receptor (6,11,43,44).In 1979, a significant human-poisoning incident was reported on Palm Island in Australia (14, 16). Afterwards, it was shown that the compound responsible for that poisoning was cylindrospermopsin (CYN) (Fig. 1) and that this cytotoxic molecule was produced by a cyanobacterium, Cylindrospermopsis raciborskii (28). Since then, it was shown that this molecule is also produced by cyanobacteria belonging to other genera such as Anabaena (38), Aphanizomenon (1, ...

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confidence: 99%

“…Several analytical techniques have been developed to assay concentrations of CYN in cultures or in the environment: an enzyme-linked immunosorbent assay (ELISA) and assays based on liquid chromatography coupled either to a UV detector (40) or to a mass spectrometer (3,12,13).…”

mentioning

confidence: 99%

Biosynthesis of Cylindrospermopsin and 7-Epicylindrospermopsin in Oscillatoria sp. Strain PCC 6506: Identification of the cyr Gene Cluster and Toxin Analysis

Mazmouz

Chapuis-Hugon

Mann

et al. 2010

Appl Environ Microbiol

123

117

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“…Even within the same water body, measured concentrations could be method dependent. Results from Bláhová et al [14][15][16][17][18][19][20][21][22][23][24] indicated that although concentrations determined by enzyme-linked immunosorbent assay (ELISA) and liquid chromatography-mass spectrometry (LC-MS) showed good quantitative agreement, concentrations determined by ELISA were systematically higher than concentrations determined by LC-MS, which was attributed to matrix effects (both in ELISA and LC-MS) and ELISA cross-reactivity with other unidentified derivatives of toxin CYL (Table 1). Table 1: Algal toxins and their environmentally relevant concentrations in selected waters of various regions; CYL = cylindrospermopsin; DA = domoic acid; GD = gymnodimine; MCs = microcystins; ND = Nodularin; OA = okadaic acid; PTX-2 = pectenotoxin-2-seco acid; STX = saxitoxin; ADDA-ELISA = ADDA ELISA Kit; LC-MS/MS = liquid chromatography-tandem mass spectrometry; cELISA = competitive enzymelinked immunosorbent assay; HPLC = high-performance liquid chromatography; HPLC-MS/MS = high-performance liquid chromatographytandem mass spectrometry; HPLC-ESI-MS = high performance liquid chromatography -electrospray ionization-mass spectrometry; LC-FD = liquid chromatography with fluorescence detection.…”

Section: Introductionmentioning

confidence: 99%

Current Techniques for Detecting and Monitoring Algal Toxins and Causative Harmful Algal Blooms

2014

J Environ Anal Chem

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The detection and monitoring techniques for algal toxins and the causative harmful algal blooms (HABs) are essential for the protection of aquatic lives, shellfish safety, drinking water quality, and public health. Toward the development of fast, easy, and reliable techniques, much progress has been made during the last decade for the qualitative and quantitative analysis of algal toxins. This review highlights the recent progress and new trends of these analytical and monitoring tools, ranging from in-situ quick screening protocols for the monitoring of algal blooms to mass spectrometric analysis of trace levels of various algal toxins and structural elucidation. Solid-phase adsorption toxin tracking (SPATT) deployed in the field for the passive sampling of algal toxins has been recently validated, and improved ELISA-based methods with lower detection limits for more toxins have become commercially available for both screening and routine monitoring purposes. Liquid chromatography-mass spectrometry with several recent mass spectrometric innovations has expanded our understanding of traditional toxins, their metabolites along with newly discovered toxins of ecological importance. Several established in vivo and in vitro bioassays will continue to be used as benchmark toxicological testing of algal toxins; however, newly emerged molecular probing techniques such as real-time quantitative polymerase chain reaction (qPCR) have extended our ability to trace algal toxins from causative organisms at the molecular level. New chemical and biological sensors, lab-on-chip and remote sensing of blooms being developed will hold promise for early warning and routine monitoring to better manage and protect our freshwater, coastal and marine resources from adverse impact by harmful algal blooms.

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“…Esta técnica permite providenciar uma informação quantitativa e qualitativa, contudo, para tal análise são necessários padrões da toxina purificada, os quais por vezes são difíceis de serem obtidos, além de necessitar de grandes volumes de amostra quando a toxina se encontra em baixas concentrações, e de passos para purificação das amostras (RIVASSEAU et al, 1999;RAPALA et al, 2002;MATHYS;SURHOLT, 2004;SANGOLKAR;MASKE;CHAKRABARTI, 2006;. A cromatografia líquida acoplada à espectrometria de massas (LC-MS) e a LC-MS/MS também são métodos muito difundidos que fornecem resultados mais precisos sobre a presença desta toxina, bem como disponibilizam dados referentes aos picos gerados pela ionização e fragmentação da molécula (CHISWELL et al, 1999;FALCONER et al, 1999;LI et al, 2001a;LI et al, 2001b;FASTNER et al, 2003;HUMPAGE et al, 2005;FROSCIO et al, 2008;RASMUSSEN et al, 2008;YLMAZ et al, 2008;BLÁHOVÁ et al, 2009;GALLO et al, 2009;PREUβEL et al, 2009;MAZMOUZ et al, 2010;MEREL et al, 2010;ANTAL et al, 2011;BALLOT et al, 2011;EVERSON et al, 2011;MAZMOUZ et al, 2011;Mc GREGOR et al, 2011). IV) Utilizar os dados gerados no sequenciamento gênomico das duas linhagens para mapeamentos e buscas por genes relacionados à síntese de metabólitos secundários, com foco principal na CYN e genes relacionados à fixação biológica do N 2 e diferenciação dos heterócitos.…”

Section: Métodos De Detecção De Cynunclassified

Análise genômica de isolados brasileiros de Cylindrospermopsis raciborskii com enfoque em cilindrospermopsina e fixação biológica de N2

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The first occurrence of the cyanobacterial alkaloid toxin cylindrospermopsin in the Czech Republic as determined by immunochemical and LC/MS methods

Cited by 98 publications

References 29 publications

Biosynthesis of Cylindrospermopsin and 7-Epicylindrospermopsin in Oscillatoria sp. Strain PCC 6506: Identification of the cyr Gene Cluster and Toxin Analysis

Biosynthesis of Cylindrospermopsin and 7-Epicylindrospermopsin in Oscillatoria sp. Strain PCC 6506: Identification of the cyr Gene Cluster and Toxin Analysis

Current Techniques for Detecting and Monitoring Algal Toxins and Causative Harmful Algal Blooms

Análise genômica de isolados brasileiros de Cylindrospermopsis raciborskii com enfoque em cilindrospermopsina e fixação biológica de N2

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