Determining the viability of Schistosoma mansoni cercariae using fluorescence assays: an application for water treatment
File(s)
Author(s)
Type
Journal Article
Abstract
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.
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.
Date Issued
2020-03-26
Date Acceptance
2020-03-11
Citation
PLOS Neglected Tropical Diseases, 2020, 14 (3)
ISSN
1935-2727
Publisher
Public Library of Science
Journal / Book Title
PLOS Neglected Tropical Diseases
Volume
14
Issue
3
Copyright Statement
© 2020 Braun et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Grant Number
EP/P028519/1
Subjects
Science & Technology
Life Sciences & Biomedicine
Infectious Diseases
Parasitology
Tropical Medicine
IN-VITRO
INHIBITION
VIVO
Animals
Cercaria
Disease Transmission, Infectious
Microscopy, Fluorescence
Parasite Load
Schistosoma mansoni
Schistosomiasis mansoni
Staining and Labeling
Treatment Outcome
Water
Water Purification
Animals
Schistosoma mansoni
Schistosomiasis mansoni
Water
Microscopy, Fluorescence
Treatment Outcome
Staining and Labeling
Water Purification
Disease Transmission, Infectious
Cercaria
Parasite Load
Tropical Medicine
06 Biological Sciences
11 Medical and Health Sciences
Publication Status
Published
Article Number
ARTN e0008176