Biological invasions of aquatic macroinvertebrates are gaining interest because of their potential for significant ecological and socio-economic impacts (positive and negative). In the present study, an inventory was made of the alien macroinvertebrates occurring in Flanders (northern Belgium) based on extensive existing collections of biological samples and supplemented with our additional sampling programs. Fresh and brackish waters as well as the Belgian coastal harbours, situated at the interface of the marine environment, were investigated. Over 2,500 samples containing alien macroinvertebrates were identified to species level, which allowed us to accurately map their distribution in Flanders. Alien macroinvertebrates are widespread and abundant in many watercourses in Flanders. Four new macroinvertebrate species for Flanders were discovered: Procambarus clarkii (Girard, 1852), Echinogammarus trichiatus (Martynov, 1932), Synurella ambulans (F. Müller, 1846) and Laonome calida Capa, 2007. Fifty-two alien macroinvertebrates were encountered in fresh and slightly brackish surface waters, and 21 alien species were reported for the Belgian part of the North Sea and its adjacent estuaries. Most alien macroinvertebrates collected were crustaceans and molluscs. Alien species found in fresh and brackish water mainly originate from the Ponto-Caspian area and North America; fewer species originated from Asia and South- and East-Europe. The major pathways were probably shipping and dispersal through canals. Based on observations in neighbouring countries, several additional species are expected to arrive in the near future. Follow-up work is needed to assess the ecological and economic impacts of existing alien macroinvertebrates, and a monitoring program is needed to detect new incoming species.