Larval settlement and metamorphosis of the mussel Mytilus galloprovincialis in response to biofilms

Biofilms were allowed to develop on glass slips immersed 1.0-1.5 m below the sea surface in Tachibana Bay, Nagasaki, Japan, for different periods of time from November 2003 to January 2005. The effects of age, immersion month, dry weight, bacterial and diatom densities of these biofilms on the settlement and metamorphosis of pediveliger larvae of the mussel Mytilus galloprovincialis were investigated in the laboratory. Furthermore, biofilms were subjected to various treatments to investigate the nature of the settlement and metamorphosis cue in the biofilm. Pediveliger larvae of the mussel settled and metamorphosed in response to biofilms. Settlement and metamorphosis to the post-larval stage significantly increased with the biofilm age. In addition, the biofilm activity varied depending on the immersion month (season), e.g., for biofilms with the same age, those immersed between June and August had higher activities than those immersed between November and March. The activity of the biofilm also positively correlated with the dry weight, bacterial and diatom densities. These three quantitative parameters of the biofilm were significantly affected by the film age but were not affected by the immersion month, suggesting that other parameters (e.g., community structures, extracellular products) also affected the inductive activity of the biofilm. The fixative agents (formalin and glutaraldehyde), heat, ethanol, ultraviolet irradiation and antibiotics treatments of the biofilm resulted in significant reduction or loss of its inductive activity. The survival of bacterial cells in the treated films where activities were either reduced or lost also decreased significantly. No settlement and metamorphosis were obtained when larvae were exposed to the conditioned water of the biofilm. Thus, larvae of M. galloprovincialis settled and metamorphosed in response to a cue produced by living bacteria in the biofilm. The cue may be a bacterial extracellular product which was susceptible to the above treatments.