The occurrence of multiple mating in Drosophila melanogaster is of particular interest to evolutionary biologists, as seminal fluid has some toxic effects for females. Thus, it has been predicted that the number of matings per females should be low. We have tested this prediction with seven highly polymorphic microsatellite loci in inseminated females from a Viennese D. melanogaster population. In contrast to the predicted low number of matings and previous studies in natural populations, we identified the genotypes of four to six different males fertilizing the offspring of each female tested. Potential causes and consequences are discussed.