Non-crop habitat adjacent to crops may be important for enhancing the activity of natural enemies in crops. However, it is not always clear whether natural enemies that are active in non-crop habitats actually contribute to pest suppression in adjacent crop habitats. We hypothesised that parasitic wasps that utilise the same hosts can be segregated between crop and non-crop habitats in an agro-ecosystem. We tested this hypothesis using the light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae), in vineyards and adjacent native vegetation. We experimentally measured the parasitism rate of larval E. postvittana at six and eight sites in both vineyards and the adjacent native vegetation in two consecutive years. Wild larval Tortricidae were also collected at each experimental site to assess their diversity and related parasitoids. Parasitised hosts were then identified using a PCR-based protocol to examine the parasitoids’ host ranges. The parasitoid Therophilus unimaculatus (Turner) (Hymenoptera: Braconidae) was most active in non-crop native vegetation, whereas Dolichogenidea tasmanica (Cameron) (Hymenoptera: Braconidae) parasitised the most larvae in vineyards. Parasitism of E. postvittana by D. tasmanica was higher on grape than on plantain, which indicates that host plants influence activities in different habitat. Both species shared the same range of tortricid hosts. Overall, our results indicate the two key parasitoids that attack E. postvittana differ in their pattern of habitat use. The native vegetation adjacent to crops may not enhance the activity of some natural enemies for pest control in an agricultural ecosystem.