Physiological responses of Xenopus laevis tadpoles exposed to cyanobacterial biomass containing microcystin-LR

Abstract

Cyanobacteria are the primary biomass producers and some species synthesize remarkable amounts of secondary metabolites, the so-called cyanotoxins. Several reports deal with the most common cyanotoxins, microcystins (MCs), and their effects on fishes but only a few studies investigated a natural exposure to MCs and limited information is available concerning the further aquatic vertebrate class, amphibians. In the present study, Xenopus laevis tadpoles at stage 52 (Nieuwkoop and Faber, 1994) were exposed for 1, 3, 7, and 21 days to diets containing lyophilized cyanobacterial biomass without and with microcystin-LR (MC-LR) at concentrations of 42.8 and 187.0μg MC-LR/g diet, respectively, to determine impacts on MC-LR bioaccumulation, development, stress, and biotransformation. The fate of MC-LR present in diet and water was determined in whole body using liquid chromatography with tandem mass spectrometry detection. Effects on development were assessed by recording mortality, weight and developmental stage. In parallel, mRNA levels of hypophyseal thyroid stimulating hormone (TSH) associated with metamorphosis and of gonadotropins, luteinizing hormone and follicle stimulating hormone, triggering sexual differentiation, were assessed. Concerning stress, corticosteroid levels and mRNA expression of heat shock protein 70 (HSP70) as stress biomarkers were examined. Furthermore, mRNA expression of biotransformation enzymes of all three phases as well as biomarkers for oxidative stress were determined.Surprisingly, exposure to cyanobacterial biomass containing MC-LR supplied via diet as natural exposure neither resulted in measurable bioaccumulation of MC-LR nor affected dramatically development. Only minor to negligible physiological impacts on development, stress, and biotransformation mechanisms were found suggesting that X. laevis tadpoles seem to have some mechanisms to be able to cope quite well with diets containing lyophilized cyanobacterial biomass even with considerable amounts of MC-LR.