After cleaning and disinfection (C&D), surface contamination can still be present in the production environment of foodcompanies. Microbiological contamination on cleaned surfaces can be transferred to the manufactured food and consequentlylead to foodborne illness and early food spoilage. However, knowledge about the microbiological composition of residualcontamination after C&D and the effect of this contamination on food spoilage is lacking in various food sectors. In this study,we identified the remaining dominant microbiota on food contact surfaces after C&D in seven food companies and assessed thespoilage potential of the microbiota under laboratory conditions. The dominant microbiota on surfaces contaminated at 102CFU/100 cm2 after C&D was identified based on 16S rRNA sequences. The ability of these microorganisms to hydrolyzeproteins, lipids, and phospholipids, ferment glucose and lactose, produce hydrogen sulfide, and degrade starch and gelatin alsowas evaluated. Genera that were most abundant among the dominant microbiota on food contact surfaces after C&D werePseudomonas, Microbacterium, Stenotrophomonas, Staphylococcus, and Streptococcus. Pseudomonas spp. were identified infive of the participating food companies, and 86.8% of the isolates evaluated had spoilage potential in the laboratory tests.Microbacterium and Stenotrophomonas spp. were identified in five and six of the food companies, respectively, and all testedisolates had spoilage potential. This information will be useful for food companies in their quest to characterize surfacecontamination after C&D, to identify causes of microbiological food contamination and spoilage, and to determine the need formore thorough C&D.