The Old World screw-worm fly, Chrysomya bezziana Villeneuve, occurs in Africa, the Middle East, Malaysia, Indonesia and Papua New Guinea, but it is not found in Australia. Introduction into Australia from any of these source areas would threaten the viability of much of the grazing industry in the northern part of the continent. Proposed control by the sterile insect release method (SIRM) would be compromised by the existence of sibling species within C. bezziana. This study examines the degree of genetic differentiation throughout the extensive range of the fly to assess if the degree of geographic differentiation indicates the existence of sibling species and, allows identification of the source of any introduced flies. Electrophoretic analysis of 23 loci from samples collected in southern Africa, the Middle East, Malaysia, Indonesia, and Papua New Guinea revealed 11 polymorphic loci. Overall, populations show remarkably little divergence given the geographic distribution of sample sites. None of the populations sampled were fixed for alternative electromorphs. There is no evidence from this study for the presence of sibling species within C. bezziana.