Quantitative Chemical Analysis of Defensive Secretion of Megacrania tsudai (Phasmatidae) and Effect of Actinidine on its Potential Predators.

Insects avoid predation in various ways, and some use multiple ways to avoid predation. However, the effects of comprehensive avoidance methods and the differences in avoidance methods among different life stages of insects have not been sufficiently discussed. The big head stick insect Megacrania tsudai uses background matching as its primary defense and chemical defense as its secondary defense. The aims of this study were to identify and isolate the chemical components of M. tsudai using repeatable methods, determine the amount of the main chemical compound, and reveal the effect of the main chemical compound on its predators. We established a repeatable gas chromatography-mass spectrometry (GC-MS) method to identify the chemical compounds of these secretions, and identified actinidine as the main compound. Actinidine was identified by nuclear magnetic resonance (NMR), and the amount of actinidine in each instar was calculated by constructing a calibration curve using pure actinidine. Mass ratios did not drastically change among instars. Furthermore, experiments involving dropping an aqueous solution of actinidine demonstrated removal behavior in geckos, frogs, and spiders. These results indicated that M. tsudai conducts secondary defenses using defensive secretions consisting mainly of actinidine.