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Determining the viability of Schistosoma mansoni cercariae using fluorescence assays: An application for water treatment.
PLOS Neglected Tropical Diseases ( IF 3.4 ) Pub Date : 2020-03-26 , DOI: 10.1371/journal.pntd.0008176
Laura Braun 1 , Lucinda Hazell 1 , Alexander J Webb 2 , Fiona Allan 3 , Aidan M Emery 3 , Michael R Templeton 1
Affiliation  

BACKGROUND 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. METHOD 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. CONCLUSION 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.

中文翻译:


使用荧光测定法确定曼氏血吸虫尾蚴的活力:水处理的应用。



背景技术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蜗牛中间宿主排出,并穿透人类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)是减少疾病传播的一种方法;它可以杀死或清除尾蚴,为人们提供安全的水用于洗澡或洗衣等活动,以替代受感染的湖泊或河流。目前,没有标准方法来评估水处理过程对尾蚴的有效性。在显微镜下检查尾蚴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的高通量测定法来量化尾蚴活力。方法 我们使用伦敦自然历史博物馆血吸虫病收藏品中受感染的蜗牛释放的曼氏尾蚴,测试了两种荧光测定法准确测定水样中尾蚴活力的能力。这些检测由双重染色组成,即活体和非活体染料;荧光素二乙酸酯 (FDA) 和 Hoechst,以及 FDA 和碘化丙啶。我们还将荧光测定的结果与显微镜测定的活力进行了比较。结论 两种荧光检测均可检测尾蚴活力,准确度至少为 92.2% ± 6.3%。将测定法与显微镜法进行比较,发现该方法的活力结果之间没有统计学上的显着差异。 然而,与显微镜相比,荧光检测主观性更小,耗时也更少,因此为量化水样中血吸虫尾蚴的活力提供了一种有前途的方法。
更新日期:2020-03-27
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