Ecotoxicological assessment of cyclic peptides produced by a Planktothrix rubescens bloom: Impact on aquatic model organisms

Cyanobacterial blooms, a natural phenomenon in freshwater ecosystems, have increased in frequency and severity due to climate change and eutrophication. Some cyanobacteria are able to produce harmful substances called cyanotoxins. These metabolites possess different chemical structures and action mechanisms representing a serious concern for human health and the environment. The most studied cyanotoxins belong to the group of microcystins which are potent hepatotoxins. Anabaenopeptins are another class of cyclic peptides produced by certain species of cyanobacteria, including Planktothrix spp. Despite limited knowledge regarding individual effects of anabaenopeptins on freshwater organisms, reports have identified in vivo toxicity in representatives of freshwater zooplankton by cyanobacterial extracts or mixtures containing anabaenopeptins. This study focused on the isolation and toxicity evaluation of the cyanotoxins produced in the 2022 Planktothrix rubescens bloom in Averno lake, Italy. The three main cyclic peptides have been isolated and identified by nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry (MS) and optical analyses as anabaenopeptins A and B, and oscillamide Y. Ecotoxicological tests on the aquatic model organisms Daphnia magna (crustacean), Raphidocelis subcapitata (algae), and Aliivibrio fischeri (bacterium) revealed that anabaenopeptins A and B do not generate significant toxicity at environmentally relevant concentrations, being also found a stimulatory effect on R. subcapitata in the case of anabaenopeptin A. By contrast, oscillamide Y displayed toxicity. Ecological implications based on ECOSAR predictions align with experimental data. Moreover, long-term exposure bioassays on different green unicellular algae species showed that R. subcapitata was not significantly affected, while Scenedesmus obliquus and Chlorella vulgaris exhibited altered growth patterns. These results, together with the alreadyknown background in literature, highlight the complexity of interactions between organisms and the tested compounds, which may be influenced by species-specific sensitivities, physiological differences, and modes of action, possibly affected by parameters like lipophilicity.