microcystin and Diarrhea

Microcystins are produced by bloom-forming cyanobacteria and pose significant health and ecological problems. In this study we show that zebra mussels respond differently to different strains of Microcystis aeruginosa, and that a highly toxic strain causes zebra mussels to produce large quantities of mucous pseudofaeces, 'pseudodiarrhoea', that are periodically expelled hydraulically through the pedal gape by shell valve adductions rather than by the normal ciliary tracts. Analysis of the pseudofaecal ejecta showed that the proportion of Microcystis aeruginosa relative to Asterionella formosa was high in the pseudofaeces and even higher in the 'pseudodiarrhoea' when a mixed diet was given to the mussels. This confirms that very toxic Microcystis aeruginosa were preferentially being rejected by comparison with the non-toxic diatom in the pseudofaeces and even more so in the 'pseudodiarrhoea'. Such selective rejection was not observed with low or non-toxic strains and would therefore tend to enhance the presence of toxic Microcystis aeruginosa in mixed Microcystis aeruginosa cyanobacterial blooms, as well as transferring toxins from the water column to the benthos. The observed acute irritant response to the toxin represents the first demonstration of an adverse sublethal effect of microcystins on invertebrate ecophysiology. Our results also suggest that it could be a specific response to microcystin-LF, a little studied toxin variant.

Topics: Animals; Bacterial Toxins; Diarrhea; Diatoms; Diet; Dreissena; Feeding Behavior; Gastrointestinal Tract; Microcystins; Microcystis; Peptides, Cyclic

A new analytical strategy was established to improve the determination and identification performance during analyses of microcystins and diarrhetic shellfish poisoning (DSP) toxins in different matrices. Automated high performance size exclusion chromatography (gel permeation chromatography, SEC) was applied for the clean-up of raw extracts from algae and mussel tissue containing either microcystins or DSP toxins. The cleaned raw extracts are well suited for the direct determination of microcystins and DSP toxins by HPLC/MS. The analyses of cleaned raw extracts containing microcystin by HPLC and UV/diode array detection (DAD) revealed chromatograms without interfering peaks. Additionally, methods for the identification of unknown microcystins and those not available as standards were developed and established. The proposed strategy is exemplarily demonstrated for the analyses of a natural algae community from a lake in Slowakia and a naturally contaminated mussel from Portugal.

Topics: Carcinogens; Chromatography, High Pressure Liquid; Diarrhea; Enzyme Inhibitors; Isoenzymes; Marine Toxins; Microcystins; Peptides, Cyclic; Phosphoprotein Phosphatases; Spectrum Analysis

Capillary electrophoresis (CE) coupled with liquid chromatography (LC)-linked protein phosphatase (PPase) bioassay was used to detect sensitivity both diarrhetic shellfish toxins and hepatotoxic microcystins in marine and freshwater samples. This procedure provided a quantitative bioscreen for the rapid optical resolution of either of these toxin families in complex mixtures such as cultured marine phytoplankton, contaminated shellfish and cyanobacteria (natural assemblages). Following detection, identified toxins were purified by an enzyme bioassay-guided two-step LC protocol. Using the latter approach, at least four microcystins were rapidly isolated from a cyanobacteria bloom (largely Microcystis aeruginosa) collected from a Canadian drinking-water lake, including a novel microcystin termed microcystin-XR, where X is a previously unidentified hydrophobic amino acid of peptide residue molecular mass 193 Da. The unified CE/LC-linked PPase bioscreen described provides a powerful capability to dissect multiple toxin profiles in marine or freshwater samples contaminated with either okadaic acid or microcystin classes of toxin.

Topics: Animals; Biological Assay; Chromatography, Liquid; Diarrhea; Electrophoresis; Ethers, Cyclic; Fresh Water; Marine Toxins; Microcystins; Okadaic Acid; Peptides, Cyclic; Phosphoprotein Phosphatases; Seawater; Shellfish