Factors responsible for species distribution of benthic macroinvertebrates, including responses at different spatial scales, have been previously investigated. The aim of the present research was to review the most relevant factors explaining chironomid species distribution focusing on factors operating at different spatial scales, such as latitude, longitude, altitude, substrate, salinity, water temperature, current velocity, conductivity, acidity, dissolved oxygen, nutrient content etc. acting at regional levels and at a large or small water basin level. Data including chironomid species abundances from different lentic and lotic waters in Italy and other surrounding countries were analyzed using partial canonical correspondence analysis (pCCA) and multiple discriminant analysis (DISCR). Spatial analyses, including univariate Moran’s I correlograms, multivariate Mantel correlograms and Moran’s eigenvector maps (MEMs), were thereafter carried out. The results showed that habitat type, including different types of lotic waters (i.e., kryal, crenal, rhithral, potamal) and different lake types (i.e., littoral, sublittoral, profundal zones), is the most significant factor separating chironomid assemblages, while spatial factors act only as indirect influencers.