Axenic growth in vitro of ancylostoma duodenal to the filariform [resting] larvae

ABSTRACT

Large numbers of free-living stages of Ancylostoma duodenale can be grown in vitro on formalin-killed Escherichia coli supplemented with cholesterol and Dulbecco's minimum essential medium [DMEN]. Eggs at 28C hatched and released first-stage larvae in 24 hours. First-stage larvae molted in 36 hours at 28C. Second-stage larvae molted in 2-3 days at 28C, but retained the old cuticle. Third-stage larvae characterized by the presence of a buccal cap capsule, sheath, filariform esophagus and absence of lips. Lateral alae were observed in all free-living stages