Freshwater cyanobacteria produce several bioactive secondary metabolites with diverse chemical structure, which may achieve high concentrations in the aquatic medium when cyanobacterial blooms occur. Some of the compounds released by cyanobacteria have allelopathic properties, influencing the biological processes of other phytoplankton or aquatic plants. These kinds of interactions are more easily detectable under laboratory studies; however their ecological relevance is often debated. Recent research has discovered new allelopathic properties in some cyanobacteria species, new allelochemicals and elucidated some of the allelopathic mechanisms. Ecosystem-level approaches have shed some light on the factors that influence allelopathic interactions, as well as how cyanobacteria may be able to modulate their surrounding environment by means of allelochemical release. Nevertheless, the role of allelopathy in cyanobacteria ecology is still not well understood, and its clarification should benefit from carefully designed field studies, chemical characterization of allelochemicals and new methodological approaches at the "omics" level.

We investigated the intestinal uptake and adverse effects of microcystins ingested with Microcystis on Daphnia galeata. The gut structure, blood microcystin concentration, appearance, and movements of Daphnia fed Microcystis PCC 7806 or a microcystin-deficient PCC 7806 mutant were monitored over time. Microcystins were rapidly taken up from the digestive cavity into the blood. This process apparently required a preceding disruption of the gut epithelium by an as-yet-unknown Microcystis factor. Once microcystins entered the blood, they affected the neuromuscular communication or another life function that influences major muscle systems. Consequently, the beat rates of the thoracic legs, mandibles, and second antennae as well as the activity of the foregut decreased, whereas the midgut muscles were stimulated. Finally, the animals exhibited symptoms of exhaustion and died. The present results suggest that an ingestion of between 10.2 ng and 18.3 ng of microcystin per 1 mg of Daphnia body fresh weight is sufficient to kill D. galeata within 2 d.Many freshwater cyanobacteria share the ability to synthesize bioactive compounds that may affect other organisms. In particular, the bloom-forming taxa, which are members of the orders Nostocales and Chroococcales, including Microcystis, Anabaena, Aphanizomenon, and Planktothrix spp., are rich sources of potent enzyme inhibitors, cellular disrupters, and compounds with a diverse range of other biological activities (Carmichael 1992;Codd 1995). Some of these metabolites present serious threats to animal and human health. Prominent examples are the microcystins, which frequently are produced by Microcystis sp. and have the potency to induce various harmful effects, such as an inhibition of protein phosphatases (e.g., MacKintosh et al. 1990).Microcystins usually are cell-bound. Although Microcystis cells may exhibit a limited excretion of microcystins 1 Corresponding author (thomas.rohrlack@niva.no). AcknowledgmentsThe authors express their gratitude to N. Willumsen for his laboratory assistance. The authors also thank J. Weckesser, W. Lampert, and M. Henning for providing stock cultures of Microcystis, Scenedesmus, and Daphnia.

Cyanobacterial hepatotoxins (microcystins and nodularins) cause numerous animal poisonings worldwide each year and are threats to human health. However, we found that extracts from several cyanobacteria isolates failed to induce hepatotoxicity even if they contained high concentrations of the liver toxin microcystin. The antitoxic activity abolishes all morphological hallmarks of microcystin-induced apoptosis, and therefore invalidates cell-based assays of the microcystin content of bloom-forming cyanobacteria. The antitoxin was purified from a cyanobacterial isolate (Nostoc sp. XSPORK 13A) from the Baltic Sea, and the activity was shown to reside in a novel cyclic peptide of the nostocyclopeptide family (nostocyclopeptide M1, Ncp-M1) that consists of seven amino acids (Tyr1-Tyr2-D-HSe3-L-Pro4-L-Val5-(2S,4S)-4-MPr6-Tyr7; MW=881) with an imino linkage between Tyr1 and Tyr7. Ncp-M1 did not compete with labelled microcystin for binding to protein phosphatase 2A; this explains why the antitoxin did not interfere with phosphatase-based microcystin assays. Currently used agents that interfere with microcystin action, such as inhibitors of ROS formation, microcystin uptake and Cam-kinase activity, are themselves inherently toxic. Since Ncp-M1 is potent and nontoxic it promises to become a useful mechanistic tool as soon as its exact cellular target is elucidated.

Aqueous extracts and organic solvent extracts of isolated marine cyanobacteria strains were tested for antimicrobial activity against a fungus, Gram-positive and Gram-negative bacteria and for cytotoxic activity against primary rat hepatocytes and HL-60 cells. Antimicrobial activity was based on the agar diffusion assay. Cytotoxic activity was measured by apoptotic cell death scored by cell surface evaluation and nuclear morphology. A high percentage of apoptotic cells were observed for HL-60 cells when treated with cyanobacterial organic extracts. Slight apoptotic effects were observed in primary rat hepatocytes when exposed to aqueous cyanobacterial extracts. Nine cyanobacteria strains were found to have antibiotic activity against two Gram-positive bacteria, Clavibacter michiganensis subsp. insidiosum and Cellulomonas uda. No inhibitory effects were found against the fungus Candida albicans and Gram-negative bacteria. Marine Synechocystis and Synechococcus extracts induce apoptosis in eukaryotic cells and cause inhibition of Gram-positive bacteria. The different activity in different extracts suggests different compounds with different polarities.

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