Marine aggregates are densely colonized by bacteria, and inter-specific interactions such as inhibition are important for colonization by aggregate-associated bacteria and thus affect the turnover of organic matter in the sea. In order to study antagonistic activities we carried out inhibition tests with 51 isolates obtained exclusively from aggregates of the German Wadden Sea. 16S rRNA gene sequences of all isolates revealed that 35% of the isolates affiliated with the Flavobacteria/Sphingobacteria group, 24% and 16% with α- and γ-Proteobacteria, respectively, 16% with the Bacillus/Clostridium group, and 10% with Actinobacteria. The relatively high percentage of Gram-positive bacteria may be related to specific features of the Wadden Sea environment. After 11 days of incubation using Burkholder agar diffusion assays the percentage of inhibitory isolates was 54.1% and this decreased to 20.7% after 20 days of incubation but it did not decline for members of the Bacillus/Clostridium group. Inhibitory activity was expressed in strain-specific patterns even though some isolates were closely related according to their 16S rRNA gene sequences. Antagonistic activity was lowest for Flavobacteria/Sphingobacteria (35%) and highest for Actinobacteria (80%). We further examined whether growth of isolates was affected when they were placed on lawns of certain other isolates. In parallel with lowest percentage of inhibitory isolates, highest growth occurred on lawns of the Flavobacteria/Sphingobacteria group whereas it was lowest on lawns of Actinobacteria and the Bacillus/Clostridium group. The high inhibitory activity of both groups of Gram-positive bacteria fits well with data from chemical screening using matrix-assisted laser desorption ionization time of flight mass spectrometry. Hence, inhibitory activity greatly influences inter-specific interactions and may impact microbial degradation and remineralization of particulate organic matter in aquatic environments.