Ontogenetic shifts in the metabolomic profiles of Battus polydamas (Lepidoptera: Papilionidae: Troidini) Feeding on Aristolochia gigantea

Abstract

Insect development involves profound physiological reorganization that can alter the assimilation and allocation of metabolites across life stages. In herbivorous insects, these processes are tightly linked to diet-derived chemical exposure, yet how metabolic organization shifts throughout ontogeny remains poorly understood. Here, we investigated ontogenetic metabolomic variation in Battus polydamas feeding on Aristolochia gigantea, a system characterized by a specialized ecological interaction between insect and host plant. We examined how chemical profiles vary across larval, pupal, and adult stages, as well as in the host plant and larval frass, to determine whether metamorphosis is associated with stage-specific reorganization of the insect metabolome. Multivariate analyses revealed clear separation among sample types, and PERMANOVA confirmed significant differences among groups. Pupae and adult bodies showed the greatest similarity, whereas plant and frass formed distinct clusters. Wings exhibited a differentiated metabolomic profile. Molecular networking revealed a diverse set of primary and secondary metabolites, including carbohydrates, amino acids, phenolics, fatty acids, and sterol-related compounds. Nine metabolites differed significantly among plant, insect stages, and frass. Lipid- and sterol-related compounds showed stage-specific variation, particularly in pupae and wings, while phenolic derivatives and fatty acids distinguished insect tissues from plant and frass samples. Frass displayed marked chemical divergence, consistent with differential assimilation and metabolic processing. In summary, the metabolomic profile of B. polydamas is reorganized across ontogeny, reflecting stage-specific patterns of metabolite retention and transformation throughout development.