Real-Time Monitoring of Helminth Motility

The xWORM technique was employed using an xCELLigence SP system (ACEA Biosciences Inc., USA) as described previously16 45 (link). All experiments were carried out as per the manufacturer’s instructions with 15 sec read intervals using the real time cell assay (RTCA) software (ACEA Biosciences Inc., USA). Separated (single) and paired adult S. mansoni flukes per well (mixed gender) were used in the xWORM assay (two independent experiments), and four adult T. muris of mixed gender were placed in each well of the 96 well E-plates containing a final volume of 200 μl of culture medium. All assays were conducted in triplicate. Inter-well spaces were filled with 100 μL of culture medium or PBS to prevent evaporation. The E-plates containing worms were cultured overnight at 37 °C with 5% CO₂ to obtain a baseline motility reading. The worms were then treated with prepared concentrations of the test compounds and the motility of the worms was monitored for 12–40 h.