Non-indigenous species (NIS) in the Mediterranean Sea. It presents a report on the softbottom macrobenthos from the five Adriatic ports: Bari, Ancona (Italy), Koper (Slovenia), Pula, Rijeka (Croatia), with a focus on the presence and contribution of NIS to native assemblages. Out of 451 species identified, only four were common to all ports. A total of eight NIS were recorded, five in surveyed ports and three in the lagoon connected to the Port of Koper. The highest number of NIS was recorded in Bari, and the highest abundance in Ancona and Bari. Generally, the number, abundance and contribution of NIS seems too low to cause a substantial impact on native communities in surveyed ports. The suitability of methods adopted for PBS for soft-bottom NIS was discussed and suggestion for methodological improvement is provided.