Department of Civil and Environmental Engineering, Imperial College London, London, United Kingdom.
Section of Structural and Synthetic Biology, Department of Infectious Disease, Imperial College London, London, United Kingdom.
PLoS Negl Trop Dis. 2020 Mar 26;14(3):e0008176. doi: 10.1371/journal.pntd.0008176. eCollection 2020 Mar.
Schistosome cercariae are the human-infectious stage of the Schistosoma parasite. They are shed by snail intermediate hosts living in freshwater, and penetrate the skin of the human host to develop into schistosomes, resulting in schistosomiasis infection. Water treatment (e.g. filtration or chlorination) is one way of cutting disease transmission; it kills or removes cercariae to provide safe water for people to use for activities such as bathing or laundry as an alternative to infested lakes or rivers. At present, there is no standard method for assessing the effectiveness of water treatment processes on cercariae. Examining cercarial movement under a microscope is the most common method, yet it is subjective and time-consuming. Hence, there is a need to develop and verify accurate, high-throughput assays for quantifying cercarial viability.
We tested two fluorescence assays for their ability to accurately determine cercarial viability in water samples, using S. mansoni cercariae released from infected snails in the Schistosomiasis Collection at the Natural History Museum, London. These assays consist of dual stains, namely a vital and non-vital dye; fluorescein diacetate (FDA) and Hoechst, and FDA and Propidium Iodide. We also compared the results of the fluorescence assays to the viability determined by microscopy.
Both fluorescence assays can detect the viability of cercariae to an accuracy of at least 92.2% ± 6.3%. Comparing the assays to microscopy, no statistically significant difference was found between the method's viability results. However, the fluorescence assays are less subjective and less time-consuming than microscopy, and therefore present a promising method for quantifying the viability of schistosome cercariae in water samples.
血吸虫尾蚴是血吸虫寄生虫的人类感染阶段。它们由生活在淡水中的中间宿主蜗牛释放,并穿透人体宿主的皮肤发育成血吸虫,导致血吸虫病感染。水处理(例如过滤或氯化)是切断疾病传播的一种方法;它可以杀死或去除尾蚴,为人们提供用于洗澡或洗衣等活动的安全水,以替代受污染的湖泊或河流。目前,没有评估水处理过程对尾蚴效果的标准方法。在显微镜下检查尾蚴的运动是最常见的方法,但它是主观的且耗时的。因此,需要开发和验证准确、高通量的定量尾蚴活力检测方法。
我们使用伦敦自然历史博物馆血吸虫收藏库中从受感染蜗牛释放的曼氏血吸虫尾蚴,测试了两种荧光检测方法对水样中尾蚴活力的准确测定能力。这些检测方法由双重染色组成,即活细胞和非活细胞染料;荧光素二乙酸酯(FDA)和 Hoechst,以及 FDA 和碘化丙啶。我们还将荧光检测的结果与显微镜下的活力测定进行了比较。
两种荧光检测方法都能以至少 92.2%±6.3%的准确度检测尾蚴的活力。将这些检测方法与显微镜相比,发现它们的活力结果之间没有统计学上的显著差异。然而,荧光检测方法比显微镜更客观、更省时,因此是一种很有前途的定量检测水样中血吸虫尾蚴活力的方法。