The knowledge of the endemic drosophilid assemblage is a useful reference to study population dynamics when new species are introduced in a geographical area. The introduction of invasive species can change the structure of the drosophilid community; hence, the distribution data for endemic species are also essential to support efficient pest management. We provide the first description of the natural drosophilid populations (Diptera Drosophilidae) recorded in Apulia, in Southern Italy. The flies, which were collected in a field survey throughout a year, were classified by morphological and molecular analyses by sequencing the barcode fragment of the COI mtDNA gene. The identified species show a distribution of frequencies that varies throughout the year, reflecting a seasonal life cycle peculiar to each species. Among the recorded drosophilids, the potential pest species Drosophila suzukii represents one of the four most abundant species
