Fluorescent Imaging with Hoechst 34580 and Propidium Iodide in Determination of Toxic Changes of Cyanobacterial Oligopeptides in Rotifers

Authors: Adam Bownik, Małgorzata Adamczuk, Barbara Pawlik-Skowrońska

Abstract:

Certain strains of cyanobacteria, microorganisms forming water blooms, produce toxic secondary metabolites. Although various effects of cyanotoxins in aquatic animals are known, little data can be found on the influence of some cyanobacterial oligopeptides beyond microcystins. The aim of the present study was to determine the toxicity of novel pure cyanobacterial oligopeptides: microginin FR-1 (MGFR1) and anabaenopeptin-A (ANA-A) on a transparent model rotifer Brachionus calyciflorus with the use of fluorescent double staining with Hoechst 34580 and propidium iodide. The obtained results showed that both studied oligopeptides decreased the fluorescence intensity of animals stained with Hoechst 34580 in a concentration-dependent manner. On the other hand, a concentration-dependent increase of propidium iodide fluorescence was noted in the exposed rotifers. The results suggest that MGFR-1 and ANA-A should be considered as a potent toxic agent to freshwater rotifers, and fluorescent staining with Hoechst and propidium iodide may be a valuable tool for determination of toxicity of cyanobacterial oligopeptides in rotifers.

Keywords: cyanobacteria, brachionus, oligopeptides, fluorescent staining, hoechst, propidium iodide