Changes in Toxin Quantities Following Experimental Manipulation of Toxin Reserves in Bufo bufo Tadpoles

Possessing toxins can contribute to an efficient defence against various threats in nature. However, we generally know little about the energy- and time-demands of developing toxicity in animals, which determines the efficiency of chemical defence and its trade-off with other risk-induced phenotypic responses. In this study we examined how immersion into norepinephrine solution inducing the release of stored toxins, administration of mild stress mimicking predator attack or simple handling during experimental procedure affected the quantity and number of toxin compounds present in common toad (Bufo bufo) tadpoles as compared to undisturbed control individuals, and investigated how fast toxin reserves were restored. We found that total bufadienolide quantity (TBQ) significantly decreased only in the norepinephrine treatment group immediately after treatment compared to the control, but this difference disappeared after 12 h; there were no consistent differences in TBQ between treatments at later samplings. Interestingly, in the norepinephrine treatment approximately half of the compounds characterized by >700 m/z values showed the same changes in time as TBQ, but several bufadienolides characterized by <600 m/z values showed the opposite pattern: they were present in higher quantities immediately after treatment. The number of bufadienolide compounds was not affected by any treatments, but was positively related to TBQ. Our study represents the first experimental evidence that toxin quantities returned to the original level following induced toxin release within a very short period of time in common toad tadpoles and provide additional insights into the physiological background of chemical defence in this model vertebrate species. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10886-019-01045-9) contains supplementary material, which is available to authorized users.