Surface properties affect the attachment of micro- and macroscopic marine organisms. The current study examined the settlement response of the mussel Mytilus coruscus plantigrades to natural biofilms formed on surfaces of different wettability. The percentages of plantigrade settlement were not influenced by the biofilms formed on variously wettable surfaces in the short term, but after 10 days, the plantigrade settlement rates decreased on biofilms formed on lower wettability surfaces. In general, lower wettability of the surfaces resulted in the decrease of the dry weight, bacterial and diatom density and the thickness of natural biofilms when compared to high wettability surfaces. In contrast, chlorophyll-a concentration in biofilms was independent of the initial wettability of the surfaces. Comparative cluster analysis of bacterial denaturing gradient gel electrophoresis patterns revealed that high variability existed between the bacterial community on high wettability surfaces and that on low wettability surfaces. Thus, surface wettability affects the formation of natural biofilms, and this variation in biofilms developed on different wettability surfaces may explain the discrepancy in their corresponding inducing activities on M. coruscus plantigrade settlement. This finding provides new insight into interactions between mussel settlement, biofilm characteristics and surface properties.