Listeria monocytogenes is a major concern for the food industry. It is one of the major agents causing listeriosis. The objective of the first part of this study is to evaluate the effect of hydration level on attached listeria monocytogenes on stainless steel/High density polyethylene transferred to food products. Attached cells were prepared on stainless steel/High density polyethylene. Transfer experiments were conducted from inoculated surface material slides to cold smoked salmon fillets. This experiment was repeated 6 times. The results were analyzed with an analysis of variance by SAS. The differences between the different RH% and surface conditions were not statistically significant. There was variability in between packages, brands and over the course of storage after opening, and likely contributed to the variability of transfer observed in this set of experiments. The objective of the second part of the research is to study the effect of hydration level on the detachment of Listeria monocytogenes biofilm growing on stainless steel by using Atomic force microscope. Biofilms were grown on stainless steel in drip flow bioreactor at 32 °C for 72 h. Then biofilms were equilibrated over saturated salt solution at 20 °C for 48 h before the Atomic Force Microscope experiment. The results showed that cohesive energy value of the biofilm increased with biofilm depth. Only square shaped displaced 2.5X2.5 μm region were visualized after serious of raster scanning under high load which means that moisture condition of Listeria monocytogenes biofilm can significantly affect the cohesiveness between of Listeria monocytogenes biofilm.