ABSTRACT This study was designed to evaluate the acaricidal effect and the mode of action of lemongrass essential oil (LEO) against Aleuroglyphus ovatus. Contact and fumigant mortality indicated that LEO had higher acaricidal activities at a low concentration, and the acaricidal action of LEO was significantly more effective by contact effects than by fumigant bioassay. Repellent rate against A. ovatus at different concentrations of LEO increased with application time. Observation on the toxic symptoms revealed that almost all mites were in immobilized type (IM type) in 30 min, but the symptom of knock-down type (KD type) appeared later. Cuticles of mites killed by LEO exhibited obvious colour change and deformation. Detoxification enzyme activity assay showed that glutathione S-transferase (GST) activity increased in mites treated by LEO compared to the controls. LEO increased catalase (CAT) activity of A. ovatus but inhibited superoxide dismutase (SOD) after 20 h. LEO also increased nitric oxide synthetase (NOS) and inhibited acetylcholine esterase (AChE) activities. Moreover, LEO acted as a fast-acting neurotoxin on A. ovatus. Our results suggest that LEO has an acaricidal potential and is a promising agent in control of A. ovatus.