The aim of the study is to explore the mixed biofilm interaction among dominant spoilage bacteria isolated from meat. The dual-biofilm characteristics of Pseudomonas lundensis and three spoilers were evaluated using by crystal violet staining， phenol sulfuric acid assay， bicinchoninic acid assay and confocal laser scanning microscope (CLSM) observation at 15 ℃ and 25 ℃， as well as protease activity and disinfectant tolerance. The results showed that Pseudomonas spp. accounted for 43.5%， followed by Brochothrix thermosphacta， Acinetobacter spp. and Myriodes spp. among 23 spoilers isolated from beef. The amount of biofilm formation of PL， AJ and BT at 15 ℃ was significantly higher than that at 25 ℃， and levels of dual-biofilms at 15 ℃ was significantly more than single culture， indicating that low temperature significantly promoted the formation of mixed biofilm. The number of cells in biofilm co-cultured with PL and AJ was 0.75-0.78 lg(CFU/cm2)， which were higher than that of single culture. Compared to PL single biofilm， the secretion of extracellular polysaccharides increased by 15.67-22.01 μg/mL， and the production of extracellular protein co-cultured with AJ or BT rose by 7.35 μg/mL and 6.70 μg/mL respectively. CLSM observation also revealed that the dual-biofilms of PL and AJ or MP formed clusters with heterogeneous structures， and their thickness of these biofilm increased to 48.99-49.22 μm. In addition， the co-culture of PL and three strains significantly enhanced its protease activity and chemical tolerance. After treatment with 0.06% sodium hypochlorite， the residual rate of dual-biofilms of PL， AJ or BT was 95.61% and 96.16% at 15 ℃， which was significantly higher than that of PL single biofilm. Thus， it was revealed that meat-derived spoilage bacteria PL and three strains formed a complex dual-biofilm with strains-dependence， and the PL cells as dual-biofilm formed at low temperature enhanced its resistance to sodium hypochlorite. This study provided a good foundation for exploring the contamination and control of mixed biofilms from meat-derived spoilage microorganisms.