THE NUMBER OF RECENT FOODBORNE ILLNESSES ASSOCIATED WITH FRESH PRODUCE IS CONCERNING. ALTHOUGH FOOD SAFETY SYSTEMSFOR FOOD PROCESSING INDUSTRIES ARE USUALLY IN PLACE, OUTBREAKS CONTINUE. ONE POSSIBLE OVERLOOKED SCENARIO FOR WHY OR HOW FOODBORNE PATHOGENS ARE PRESENT IN INFECTIOUS DOSE LEVELS IN PACKAGED AND BULK PRODUCE MAY BE THAT ONCE PATHOGENS HAVE ENTERED THE PRODUCTION CHAIN, THEY ARE PROTECTED OR GO UNDETECTED BY BEING SEQUESTERED WITHIN PROTOZOA OR PROTOZOAN CYSTS THAT MAY BE RESISTANT TO SANITIZERS IN THE PRODUCE INDUSTRY. WE HYPOTHESIZE THAT CYST-FORMING PROTOZOA ARE PRESENT IN THE FOOD PACKAGING ENVIRONMENT AND ON PRODUCE AND MAY ENTRAP AND PROTECT FOODBORNE PATHOGENS.TO DETERMINE WHETHER THESE PROTOZOA ARE IMPORTANT IN THE PROTECTION OF FOODBORNE PATHOGENS, WE WILL 1) ISOLATE AND CULTURE CYST-FORMING PROTOZOA FROM PRODUCE AND TEST ENTRAPMENT OF THE FOODBORNE PATHOGENS, E. COLI O157:H7, LISTERIA, AND SALMONELLA, IN CYSTS OF AMOEBAE AND CILIATES. WE WILL THEN USE THE CONDITIONS FOUND IN PRODUCE PACKAGING PLANTS TO TEST RELEASE OF VIABLE FOODBORNE PATHOGENS FROM PROTOZOAN CYSTS. THE CONDITIONS USED WILL INCLUDE EXPOSURE TO SANITIZERS AND MODIFIED ATMOSPHERE PACKAGING (CONDITIONS IN THE BAGS OF PRE-PACKAGED LEAFY GREENS FOUND IN GROCERY STORES). WE WILL ALSO TEST WHETHER SANITIZERS AND MODIFIED ATMOSPHERE PACKAGING CAN INDUCE PROTOZOA TO FORM CYSTS AND TRAP PATHOGENS. LASTLY, WE WILL CHARACTERIZE THE PROTOZOAN MICROBIOME OF FRESH PRODUCE AND IN PACKAGING WASH WATER TO DETERMINE THE DIVERSITY OF PROTOZOA THAT MIGHT BE PRESENT ON THE PRODUCEAND COULDENTRAP AND PROTECT BACTERIA. THIS PROJECTWILL HELP TO DETERMINE THE SIGNIFICANCE OF AND ROLES FOR PROTOZOA IN OUTBREAKS ASSOCIATED WITH PACKAGED PRODUCE, SUCH THAT UNDERSTANDING THE ROLES FOR THESE ORGANISMS MAY LEAD TO NEWINTERVENTIONS FOR REDUCING PROTOZOA ON PRODUCE, THEREBY REDUCING THE RISK FOR FOODBORNE ILLNESS.