Optimising the xWORM Assay for Monitoring Hookworm Larvae Motility

Parasitic worms (helminths) infect almost all taxa across the animal kingdom, and pose significant challenges to public health and economies, particularly in developing countries. To address this problem, researchers have developed various tools to measure the motility and viability of helminths. However, the conditions used in anthelmintic screening assays are often not optimised, and they can vary considerably between research teams. These un-optimised conditions can impact new drug screens as little is known about the effect of different conditions on the health of the target parasites. To improve future research this study determined the effect of key assay parameters such as media type, media concentration, in-well parasite density, and assay duration on the infective third-stage larva (L3) of two types of hookworms, <em>Nippostrongylus brasiliensis</em> in rodents and <em>Necator americanus </em>in humans. Conditions were screened over several days using the xWORM assay, a real time impedance-based helminth motility assay using the xCELLigence system with 96-well microplates. While results varied depending on the species and media used, the study found that 500-1000 L3/200µL well volume and a media concentration of 3.13 – 25% generally produced good to excellent assay conditions. This data set provides the outcomes for all conditions tested by this study. This study’s findings can guide future selection of xWORM assay parameters for novel drug trials involving these parasite species and serve as a suggested model for optimising trial conditions for alternate parasite targets and assays.寄生蠕虫（helminths）几乎可侵染动物界的所有类群，对公共卫生与经济发展造成严重威胁，在发展中国家尤为突出。为解决这一难题，科研人员已开发出多种工具用于检测寄生蠕虫的运动能力与存活活力。然而，驱虫药筛选实验中所使用的实验条件往往未经过优化，且不同研究团队间的实验条件差异显著。由于目前对不同实验条件对靶标寄生虫健康状态的影响知之甚少，这些未优化的条件可能会对新药筛选工作造成干扰。为优化后续研究工作，本研究以两类钩虫的感染性三期幼虫（L3）为研究对象——即啮齿类动物来源的巴西圆线虫（*Nippostrongylus brasiliensis*）与人类来源的美洲板口线虫（*Necator americanus*）——探究了培养基类型、培养基浓度、孔内寄生虫密度、实验时长等关键实验参数的影响。本研究通过xWORM实验（xWORM assay）——一种基于xCELLigence系统搭配96孔微孔板的实时阻抗法寄生蠕虫运动能力检测实验——连续数日对实验条件进行了筛选。尽管实验结果因所使用的寄生虫物种与培养基类型而异，但研究发现，当每孔接种500~1000条L3幼虫、总体积为200μL，且培养基浓度为3.13%~25%时，通常可获得良好至极佳的实验效果。本数据集收录了本研究中所有测试实验条件的检测结果。本研究的发现可为后续针对上述寄生虫物种开展的新药实验中xWORM实验参数的选择提供参考，同时也可为优化其他靶标寄生虫及相关实验的实验条件提供参考模型。