Sequestration of phorbol esters by aposematic larvae of Hyles euphorbiae (Lepidoptera: Sphingidae)?

Summary. The larvae of the hawkmoth species Hyles euphorbiae have a conspicuous aposematic colouration and show gregarious behaviour. It has thus been suggested that they sequester phorbol esters from their food plants which include different species of the genus Euphorbia (Euphorbiaceae) for chemical protection against predators. To test this hypothesis in more detail, we fed larvae an artificial diet with three doses of 12-tetradecanoyl-phorbol-13-acetate (TPA), then examined the faeces and the larval tissues, such as integument, haemolymph and gut of the caterpillars for the presence of TPA. In order to determine the ability of the larvae to detoxify phorbol esters, other larvae were directly injected with a TPA solution and analysed in the same manner. Our study indicates that the larvae of Hyles euphorbiae do not sequester phorbol esters. Upon oral application TPA was not found in the larval integument or the haemolymph. Instead, it was mostly metabolised (about 70–90%). Nevertheless, about 10-30% were retained and recovered in the faeces. The larvae were also able to metabolise and thus detoxify the phorbol ester when TPA was injected directly into the body. These hawkmoth caterpillars are relatively large and have a gut full of plant material, which they regurgitate into the direction of the predator when attacked in nature. Since phorbol esters are very potent toxins and irritants, we postulate that the gut content (and especially the plant slurry disgorged as regurgitant from the anterior gut) alone could be aversive for a potential predator, even if some metabolism has taken place. Thus, although H. euphorbiae caterpillars do not actively sequester phorbol esters, their aposematic colouration appears to be based on chemical defence through phorbol esters retained in the gut.