Two-day-old baby hamsters were infected initially with the infective larvae of hamster-adapted human hookworm, Necator americanus (NaL3). After a specified period they were again infected orally with infective larvae of Ancylostoma ceylanicum (AcL3). Three weeks after the second infection they were killed and the establishment of N. americanus and A. ceylanicum was assessed. The effect of different infection levels and exposure period of N. americanus on the concurrent establishment of A. ceylanicum was also examined. An infection with 50 NaL3 percutaneously, and 3 weeks later, a second infection with 50 AcL3 orally has produced reasonably equal number of hookworms (no statistical difference in the burden of N. americanus and A. ceylanicum) in the intestine of hamsters. Thus this protocol of dual infection was found suitable to develop two species of hookworms in hamsters for anthelmintic screening.