Many flies (order Diptera) act as biological control agents of pest insects and/or as pollinators of crops, and their role in ecosystems is increasingly recognized. Traditionally, the focus of biological control and pollination has been on a restricted number of species, mainly hymenopterans, such as parasitic wasps and bumblebees. However, there exists an enormous natural diversity of insect species, in particular dipterans, that may be used for such services as well. Few efforts are being directed towards developing these alternative species for exploitation and application in pest control and crop pollination. The aim of this Special Issue is to contribute to filling the knowledge gaps about the biology, production, exploitation and application of dipterans that are useful for biological control and pollination, often playing both ecosystem services. The aim is to cover both fundamental studies on the biology and ecosystem role of these flies as well as applied research into their rearing and application. The Special Issue is edited by Maria Luisa Dindo (Alma Mater Studiorum Università di Bologna, Italy), Fernanda Cingolani (National University of La Plata, Argentina) and Leo W Beukeboom (University of Groningen, The Netherlands).