Viability of developmental stages of Schistosoma mansoni quantified with xCELLigence worm real-time motility assay (xWORM).
Infection with helminth parasites causes morbidity and mortality in billions of people and livestock worldwide. Where anthelmintic drugs are available, drug resistance is a major problem in livestock parasites, and a looming threat to public health. Monitoring the efficacy of these medicines and screening for new drugs has been hindered by the lack of objective, high-throughput approaches. Several cell monitoring technologies have been adapted for parasitic worms, including video-, fluorescence-, metabolism enzyme- and impedance-based tools that minimize the screening bottleneck. Using the xCELLigence impedance-based system we previously developed a motility-viability assay that is applicable for a range of helminth parasites. Here we have improved substantially the assay by using diverse frequency settings, and have named it the xCELLigence worm real-time motility assay (xWORM). By utilizing strictly standardized mean difference analysis we compared the xWORM output measured with 10, 25 and 50 kHz frequencies to quantify the motility of schistosome adults (human blood flukes) and hatching of schistosome eggs. Furthermore, we have described a novel application of xWORM to monitor movement of schistosome cercariae, the developmental stage that is infectious to humans. For all three stages, 25 kHz was either optimal or near-optimal for monitoring and quantifying schistosome motility. These improvements in methodology sensitivity should enhance the capacity to screen small compound libraries for new drugs both for schistosomes and other helminth pathogens at large.
Department of Microbiology, Immunology & Tropical Medicine, and Research Center for the Neglected Diseases of Poverty, School of Medicine and Health Sciences, George Washington University, Washington, D.C. 20037, USA.
Gabriel
G
Rinaldi
Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, McGregor Rd, Smithfield, Cairns, Queensland 4878, Australia.
Alex
A
Loukas
Department of Microbiology, Immunology & Tropical Medicine, and Research Center for the Neglected Diseases of Poverty, School of Medicine and Health Sciences, George Washington University, Washington, D.C. 20037, USA.
Paul J
PJ
Brindley
ACEA Biosciences, San Diego, CA, USA.
Jeff T
JT
Irelan
Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, McGregor Rd, Smithfield, Cairns, Queensland 4878, Australia.
Michael J
MJ
Smout
D000818:Animals; D001681:Biological Assay; D001700:Biomphalaria; D005260:Female; D008297:Male; D051379:Mice; D009043:Motor Activity / Q000502:physiology*; D010063:Ovum; D012550:Schistosoma mansoni / Q000502:physiology*
Helminths; High-throughput; Schistosome; Strictly standardized mean difference (SSMD); Viability; xCELLigence