Bacteria were isolated and identified from the digestive tract of the secondary screwworm, Cochliomyia macellaria (F.) (Diptera: Calliphoridae), and their role in the larval development of this insect was assessed in laboratory bioassays. The analysis of 16S rDNA sequences revealed that the bacterial isolates represented four species: Providencia sp., Escherichia coli O157:H7 (Escherich), Enterococcus faecalis (Orla-Jensen), and Ochrobactrum sp. (Holmes). Developmental assays demonstrated that C. macellaria larvae are able to develop on a sterile blood agar, and no bacteria are required to complete larval development. Indeed, developmental times were shorter and survival rates of C. macellaria on a sterile blood agar and the modified Harris rearing diet were greater compared with that on the blood agar inoculated with individual and mixed bacterial isolates. The cultures of Ochrobactrum sp. and E. faecalis supported larval development to a significantly greater extent than those of Providencia sp. and E. coli O157:H7. The presence of bacteria in newly emerged C. macellaria adults also was assessed and revealed that the bacteria in the gut of larvae can survive pupation and colonize the gut of adult flies. This study shows that development of larvae of C. macellaria does not depend on bacteria and that some bacterial isolates negatively impact larval development.