A comparative study revealed hyperspectral imaging as a potential standardized tool for the analysis of cuticle tanning over insect development.

Cereal-feeding beetles are a major risk for cereal crop maintenance. Cereal weevils such as have symbiotic intracellular bacteria that provide essential aromatic amino acid to the host for the biosynthesis of their cuticle building blocks. Their cuticle is an important protective barrier against biotic and abiotic stresses, providing high resistance from insecticides. Quantitative optical methods specialized in insect cuticle analysis exist, but their scope of use and the repeatability of the results remain limited. Here, we investigated the potential of Hyperspectral Imaging (HSI) as a standardized cuticle analysis tool. Based on HSI, we acquired time series of average reflectance profiles from 400 to 1000 nm from symbiotic (with bacteria) and aposymbiotic (without bacteria) cereal weevils exposed to different nutritional stresses. We assessed the phenotypic changes of weevils under different diets throughout their development and demonstrated the agreement of the results between the HSI method and the classically used Red-Green-Blue analysis. Then, we compared the use of both technologies in laboratory conditions and highlighted the assets of HSI to develop a simple, automated, and standardized analysis tool. This is the first study showing the reliability and feasibility of HSI for a standardized analysis of insect cuticle changes.