Identification of growth inhibiting compounds in a Giardia lamblia high-throughput screen.
Giardia lamblia is one of the most common eukaryotic pathogens and is classified by the CDC as a category B agent of bioterrorism. In a departure from more traditional research focused on specific pathways or molecules, we have developed a high-throughput assay for screening libraries of small compounds for inhibitors and enhancers of trophozoite multiplication. Following a 24-h period of culture in 384-well plates in the presence of compounds, trophozoites were fixed, stained and enumerated. Quadruplicate screening of 1520 compounds from two libraries of known bioactives detected numerous inhibitory compounds. Based on a stringent cut-off of 5 standard deviations from the plate mean, 50 compounds (3.3%) were inhibitory. The activity of 3 compounds was confirmed in conventional culture. Although not meeting the threshold, one compound (indirubin) was identified as an agonist of trophozoite proliferation. Demonstrating the potential of high-throughput screening for rapidly finding new compounds which perturb G. lamblia multiplication, most of the hits identified by high-throughput screening do not appear to have been tested previously for their ability to affect G. lamblia trophozoites. High-throughput screening of bioactive compounds will open new avenues to a system-wide analysis of pathways affecting G. lamblia proliferation, and eventually to other phases of the life cycle.
Bonilla-Santiago
Rubén
R
Tufts Cummings School of Veterinary Medicine, 200 Westboro Road, North Grafton, MA 01536, USA.
Wu
Zhijin
Z
Zhang
Linghui
L
Widmer
Giovanni
G
D000818:Animals; D001681:Biological Assay / Q000379:methods; D016829:Giardia lamblia / Q000187:drug effects / Q000254:growth & development*; D006131:Growth Inhibitors / Q000737:chemistry*; D006403:Hematologic Tests; D053058:Trophozoites / Q000187:drug effects
D006131:Growth Inhibitors
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