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Simple Summary: The gut microbiota—the community of microorganisms that colonize the gut—is now recognized as a key regulator of immune activity, metabolism, and welfare in all vertebrates, including poultry. The diet and environment can both influence the gut microbiota, but the extent of these changes is unclear in poultry, where diets and environments are important management tools. As the majority of U.S. egg production (>90%) has pledged to move to cage-free egg production by 2025, it is necessary to understand how much the diet and the rearing environment contribute to gut microbiota composition and function, and ultimately to health and production traits of chicken. We addressed this unknown by analyzing the gut microbiota community of laying hens with both the housing environment and diet as variables. We compared conventional cage systems against cage-free systems. In both environments, hens were fed a standard soy-based diet, versus an alternate soy-free diet. We found that cage-free environments generated higher gut microbiota diversity, and that the diet had a relatively lower effect on changing the gut microbiota. Our results highlight the difficulty of promoting consistent, beneficial gut microbiota across production systems or diet variations in commercial poultry conditions. The gut microbiota of chicken has received much attention due to its importance for bird health, food safety, and performance. In the United States, the impending transition to cage-free housing environments has raised many questions about its consequences for poultry health, productivity, and welfare. Therefore, we investigated how housing environments and feed composition affect the poultry gut microbiome. Such data is necessary to inform the design of production systems that promote health and food safety. In this study, we investigated the cecal microbiome of both caged and cage-free laying hens that were fed either an industry-standard soy-based versus a soy-free diet. Caged hens were housed in standard industry-style layer cages with one bird per cage, and cage-free hens were housed in a poultry barn with an outdoor enclosed yard with multiple hens per pen. Our study showed significant differences in the gut microbiota between cage-free and caged environments. Cage free housing generated higher diversity compared to caged housing. Furthermore, we observed a synergistic interaction of soy-based feed in cage-free housing, as the cage-free soy group showed the highest alpha diversity, whereas the caged-soy group showed the lowest diversity overall. [ABSTRACT FROM AUTHOR]