Listeria monocytogenes (LM) is a pathogen that is found in the natural environment which can cross-contaminate produce. There is a need for greater information on the prevalence and distribution of LM along the produce supply chain and within fresh produce operations. The produce industry also needs a way to rapidly test scenarios to aid in decision-making. The ultimate objective of this study is to address these needs relevant to produce operations and provide information that will aid in the prioritization of LM control strategies. Environmental sampling plans for Listeria were implemented in eight U.S. produce operations. A total of 2,014 sponge samples were collected over one year. Isolates from this and other studies of produce operations were combined to create an isolate collection of LM (n=169) and other Listeria spp. (n=107), which were characterized using WGS. The overall findings were used to inform the development of two agent-based models of fresh-cut produce facilities that simulate, in silico, how Listeria enters, moves throughout, and leaves the built environment. These models were used to evaluate sampling schemes on their ability to locate presence of Listeria in a facility. LM prevalence varied from <0.4% to 5.8% among the sampled produce operations. LM from these and other produce operations showed virulence potential as a low proportion of LM isolates (9/169) had inlA premature stop codons, while a large proportion (83/169) had either or both of the hypervirulent-associated LIPI-3 and LIPI-4 operons. Isolates showed persistence within operations, with re-isolation (at least 2 months apart) of highly related strains (<10 hqSNP differences) found in 7/16 operations. Developed models of fresh-cut facilities evaluated sampling schemes on how well the Listeria prevalence of the collected samples reflected the true prevalence. Sampling performance of various schemes differed from each other and over time. Overall, the data shows the prevalence, distribution and virulence potential of LM isolates collected from produce operations and shows instances where closely related Listeria were found in some of these operations over time. Developed models that incorporated these findings allow for rapid virtual experimentation and evaluation of a variety of sampling schemes.