As a stage of life cycle, larval settlement and metamorphosis are critical processes for persistence of many marine invertebrate populations. Bacterial biofilms (BFs) could induce larval settlement and metamorphosis. Pseudoalteromonas, a widely distributed genus of marine bacteria, showed inductive effects on several invertebrates. However, how Pseudoalteromonas BFs induce settlement and metamorphosis of Mytilus coruscus remains unclear. Pseudoalteromonas marina BFs with the highest inducing activity were further investigated to define inductive cues. Surface-bound products of P. marina BFs could induce larval settlement and metamorphosis. P. marina BFs treated with formalin, antibiotics, ultraviolet irradiation, heat and ethanol significantly reduced inductive effects and cell survival rates. The confocal laser scanning microscopy and the biovolume analysis showed the dominance of α-polysaccharides on P. marina BFs. Treatment of BFs with amylases, proteases and lipase led to the decrease of inducing activity, suggesting that inductive cues of P. marina BFs may comprise of molecular domains of polysaccharides, proteins, and lipids. Finding inductive cues of BFs could put forward further studies about the mechanism of larval settlement and metamorphosis of marine invertebrates. Most marine benthic invertebrates have a planktonic larval phase in their life cycle, which plays an important role in their survival and development 1,2. Competent larvae choose an acceptable substratum to settle and metamorphose to the benthic stage 3,4. This critical step is affected by abiotic factors (exogenous physical and chemical) and biotic factors (endogenous and exogenous) 2,4-6. Natural biofilms (BFs) which developed in the sea have a complex structure and are composed of many species of microorganisms 7. Natural BFs were proposed as one of biotic cues to stimulate larval settlement and metamorphosis 8-10. In benthic communities, bacteria exist in a form of a bacterial BF, which is a prevalent microbial lifestyle 11,12. Not only multispecies BFs but also monospecies BFs of bacteria can induce larval settlement and metamorphosis 13,14. It has been shown that BFs of Pseudoalteromonas, Vibrio, Shewanella and Tenacibaculum can induce larval settlement and metamorphosis 3,14-16. Pseudoalteromonas species are widely distributed at various marine environments and have high ecological significance 17,18. BFs of some Pseudoalteromonas species were reported as inducers 14,19,20 or inhibitors 21-24 of larval settlement. However, it is unclear whether varying species of Pseudoalteromonas show inducing or inhibiting activity to the mussel Mytilus coruscus. Bacteria in BFs were buried in extracellular polymeric substances (EPS) which are mostly secreted by bacterial strain itself 25. Biofilm's EPS include fatty acids, proteins, polysaccharides, nucleic acids and other biopolymers 25. EPS could be divided into the water-soluble part and water-insoluble part 25-28. The water-soluble and-insoluble cues were involved in larval settlement and meta...