Little is known about the microbial communities found in distribution centers (DCs), especially in those storing and handling food. As many foodborne bacteria are known to establish residence in food facilities, it is reasonable to assume that DCs handling foods are also susceptible to pathogen colonization. To investigate the microbial communities within DCs, 16S amplicon sequencing was completed on 317 environmental surface sponge swabs collected in DCs (  = 18) across the United States. An additional 317 swabs were collected in parallel to determine if any viable species were also present at each sampling site. There were significant differences in median diversity measures (observed, Shannon, and Chao1) across individual DCs, and top genera across all reads were _A, , _E, , and based on taxonomic classifications using the Genome Taxonomy Database. Of the 39 16S samples containing ASVs, four of these samples had corresponding positive microbiological samples. Data indicated a predominance of ASVs identified as cold-tolerant bacteria in environmental samples collected in DCs. Differential abundance analysis identified , , and present at a significantly greater abundance in positive microbiological compared to those negative for . Additionally, microbiome composition varied significantly across groupings within variables (e.g., DC, season, general sampling location).