- Two-spotted spider mites (TSSM; Tetranychus urticae Koch) cause significant damage to crops and yields, in the field as well as in greenhouses. By feeding, TSSM destroy chloroplast-containing cells; this damage can be spectrally detected in the reflectance of the visible and near-infrared regions. This study focuses on hyperspectral reflectance data of greenhouse pepper (Capsicum annuum) leaves, obtained by integrated sphere. The reflectance data were transformed into vegetation indices allowing early TSSM damage detection by separation between leaf damage levels. One-way analysis of variance of coupled damage levels was applied to each of the vegetation indices. We concluded that early identification of TSSM greenhouse pepper leaf damage can be obtained by multispectral means. Furthermore, the proposed methods may identify the damage on the upper side of the leaves although the TSSM feed on the underside of leaves.