**AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** THE GOAL OF THE PROJECT IS TO DESIGN EFFECTIVE STRATEGIES FOR CONTROLLING THE DISPERSAL OF MULTI-SPECIES FOOD PATHOGENS, BASED ON ADVANCED UNDERSTANDING OF MICROBIAL INTERACTIONS AND BIOPHYSICAL FACTORS OF CONTAMINATION DYNAMICS. FOOD-BORNE PATHOGENS SUCH ASESCHERICHIA COLI,SALMONELLA ENTERICAANDLISTERIA MONOCYTOGENESFORM HIGHLYRESILIENTMULTI-SPECIES BIOFILMS IN FOOD PROCESSING FACILITIES. THEY ARE FOUND IN MANY AREAS SUCH AS THE BACK OF CONVEYOR BELTS OR DRAINS, WHICH HAVE POOR ACCESSIBILITY FOR SANITATION AND THEREFORE RESULT IN BIOFILMS WITH ENHANCED SANITIZER TOLERANCE AND ANTIMICROBIAL RESISTANCE. WE AIM TO CONTROL FOOD CONTAMINATION BY INVESTIGATING THE BIOFILM HOTSPOTS. THE SPECIFIC OBJECTIVES OF THIS PROJECT ARE TO (1) INVESTIGATE SANITIZER TOLERANCE IN FOOD PATHOGEN-MIXED-SPECIES BIOFILM (2) MEASURE THE EFFECT OF SPATIOTEMPORAL MICROSCALE PATTERNS ON BIOFILM DISPERSAL IN DRAINS OF FOOD PROCESSING SYSTEMS (3) EVALUATE BIOPHYSICAL FACTORS GOVERNING BIOFILM DETACHMENT AND DISPERSAL, LEADING TO HOTSPOT SPECIFIC INTERVENTION TO PREVENT DISPERSAL OF PATHOGENS FROM THE HOTSPOTS. THESE OBJECTIVES WILL BE ADDRESSED EXPERIMENTALLY USING COMBINATION OF OMICS, MICROSCOPIC AND MICROFLUIDIC TECHNIQUES.THE COMPREHENSIVE UNDERSTANDING OF FOOD-BORNE PATHOGEN DISPERSAL PROCESSES FROM HOTSPOTS IN FOOD PROCESSING FACILITIES AND THEIR UNDERLYING MICROBIOLOGICAL MECHANISMS, IS HIGHLY SIGNIFICANT, AS IT WILL LEAD TO POWERFUL NEW APPROACHES THAT WILL REDUCE THE CONTAMINATION OF FOOD WITH PATHOGENS.