In vitro excystation and subsequent axenic growth of Giardia lamblia

Abstract

Six fresh Giardia lamblia strains were isolated by in vitro excystation and subsequent culturing of excysted parasites in bile-supplemented BI-S-33 medium. The cysts passed in faeces appeared to differ in structure when observed using differential interference contrast microscopy. Sometimes the enclosed organisms were closely applied to the cyst wall; in most stool specimens, however, the parasites were separated from the cyst wall by a space. Cysts with parasites closely applied to the wall were the most viable type, with excystation rates up to 90%. Cysts with organisms detached from the wall displayed low excystation rates, 0-15%. During exposure to the induction solution of hydrochloric acid (pH 2), parasites initially closely applied to the cyst walls completely detached, and after transfer into culture medium vigorous flexing movements of the organisms were observed. Stool samples from 42 symptomatic and asymptomatic giardiasis patients were examined; in 26 of the samples parasites hatched, and 6 new strains were established in axenic culture.