This study aims to characterize the biofilm produced by mono- and dual-species of Shewanella baltica and Pseudomonas fluorescens as fish spoilers at the different incubation temperature, and to elucidate the interactive behaviour of dual-species biofilm development.The mono- and dual-species biofilm formation and adhesion characteristics of S. baltica and P. fluorescens were evaluated by using crystal violet staining, scanning electron microscopy and confocal laser scanning microscopy. Results showed that P. fluorescens had significantly higher biofilm biomass and polysaccharides production than S. baltica, and two isolates reached the maximum biofilm biomass during the early stationary phase. Lower biomass and polysaccharides in dual-species biofilms were observed compared to mono-species of P. fluorescens. Meanwhile, S. baltica and P. fluorescens formed fragile and viscous pellicles with different spatial architectures respectively. In dual-species pellicle few large microcolonies were dominated by P. fluorescens. Compared to mono-species of PF07, adherent cell population and biofilm thickness at the developing phase significantly decreased, and biofilm-forming cycle prolonged in the dual-species biofilms. Biofilm formation and adhesion of mono- and dual-species at 4 or 15°C were significantly higher than at 30°C during the same phase. The culture supernatant extracts of the two spoilage strains greatly inhibited biofilm development to each other.Shewanella baltica and P. fluorescens had different biofilm and pellicle characteristics, and the inhibitory development on dual-species biofilm was associated with the competitive interaction by the two psychrotrophic spoilage bacteria.This work contributes to a better understanding of interactive behaviour of multispecies biofilm communities by psychrotrophic spoilage bacteria at low temperature, which could contribute to further control contamination of spoilage organism during the preservation and processing of aquatic products.