1.Perennial bioenergy systems, such as switchgrass and restored prairies, are alternatives to commonly used annual monocultures such as maize. Perennial systems require lower chemical input, provide greater ecosystem services such as carbon storage, greenhouse gas mitigation, and support greater biodiversity of beneficial insects. However, biomass harvest will be necessary in managing these perennial systems for bioenergy production, and it is unclear how repeated harvesting might affect ecosystem services. 2.In this study, we examined how repeated production-scale harvesting of diverse perennial grasslands influences vegetation structure, natural enemy communities (arthropod predators and parasitoids), and natural biocontrol services in two states (Wisconsin and Michigan, USA) over multiple years. 3.We found that repeated biomass harvest reduced litter biomass and increased bare ground cover. Some natural enemy groups, such as ground-dwelling arthropods, decreased in abundance with harvest whereas others, such as foliar-dwelling arthropods increased in abundance. The disparity in responses is likely due to how different taxonomic groups utilize vegetation and differences in dispersal abilities. 4.At the community level, biomass harvest altered community composition, increased total arthropod abundance, and decreased evenness but did not influence species richness, diversity, or biocontrol services. Harvest effects varied with time, diminishing in strength both within the season (for total abundance and evenness), across seasons (for evenness), or were consistent throughout the duration of the study (for community composition). Greater functional redundancy and compensatory responses of the different taxonomic groups may have buffered against the potentially negative effects of harvest on biocontrol services. 5.Synthesis and applications. Our results show that in the short-term, repeated harvesting of perennial grasslands (when insect activity is low) consistently altered vegetation structure but had mixed effects on natural enemy communities and no discernable effects on biocontrol services. However, the long-term effects of repeated harvesting on vegetation structure, natural enemies, and other arthropod-derived ecosystem services such as pollination and decomposition remain largely unknown.