Raw milk and farm environment samples were collected from six farms in three regions of China for the isolation and characterization of Pseudomonas fluorescens. The biofilm-forming ability of P. fluorescens was evaluated for the screening of representative strains to investigate environmental adaptability under different culture temperatures, conditions, and surfaces. A total of 25 P. fluorescens strains were isolated and divided into five sequence types (STs). In particular, P. fluorescens 18 exhibited a more stable biofilm-forming ability at 24 h and 36 h, with a significantly higher OD value than those of other strains. P. fluorescens 18 was capable of biofilm production in tryptic soy broth, skim milk, and whole milk at 25 ℃ , 15 ℃ , and 4 ℃ , albeit with different production capacities. At a culture time of 36 h, a low temperature inhibited the formation of extracellular substances in the P. fluorescens 18 biofilm, while milk-based culture media promoted their formation. Scanning electron microscopy results showed that the adhesion ability of the P. fluorescens 18 biofilm differed among surfaces and followed the order of stainless steel>glass>plastic. These results demonstrated the risk of biofilm formation in milk-based media by wild-type P. fluorescens isolated from raw milk and its farm environment during production, with a higher risk being on stainless steel than on other surfaces. The present study provides a scientific basis for future research on the hazards of wild-type P. fluorescens.