Listeria monocytogenes is an important foodborne human pathogen associated with high mortality rates. Even if good manufacturing processes are followed, contamination can occur after processing making Listeria a major concern for food industry. As part of the EcoSec ANR program whose overall purpose is to reduce the environmental impact of hygiene operations in workshops producing chilled food, this work is more specifically focused on the adaptive response of L. monocytogenes to a desiccation stress. For that, first aim of this study was to propose a standard protocol optimized for the growth of cells in biofilm and the application of a desiccation stress that could be compared with the conditions encountered in the industry. Viabilty of bacteria measured with classical plate count method and confocal microscopy revealed clear viability differences between desiccated and control biofilm. Identification of intracellular and surface proteins involved in bacterial desiccation tolerance was investigated using different proteomic approaches.