Comparison of different staining methods for determination of viability on Mesocestoides vogae tetrathyridia
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Mesocestoides vogae is widely employed as a model for studying the biology, differentiation, and experimental chemotherapy of cestodes. Currently, there are few techniques to measure the viability of M. vogae metacestodes during pharmacological experiments. The aim of the present work was to evaluate and compare different staining techniques to determine objectively the viability of M. vogae tetrathyridia. Eosin (0.05% w/v), methylene blue (0.01% w/v), propidium iodide (PI, 2 μg/ml), and fluorescein diacetate (FDA, 0.5 μg/ml) solutions were tested against live, heat-killed (cultivated at 65 °C for 2 h) and thymol-treated tetrathyridia (50 and 250 μg/ml). Parasites were counted under a dissecting microscope or a fluorescence compound microscope, as appropriate. Studies by scanning electron microscope were performed to compare the ultrastructural damage with the viability of parasites. After comparing the performance of different dyes, we chose the eosin staining technique because its simplicity, rapidity, sensitivity, low cost and fidelity.
KeywordsMesocestoides vogae Viability Eosin Methylene blue Propidium iodide Fluorescein diacetate
The authors gratefully acknowledge the Instituto de Investigaciones Biológicas (IIB) CONICET—UNMdP for allowing us to use the fluorescence microscope. We also would like to thank Dr. Alejandra Goya (SENASA, Argentina) and Dr. Andrea Chisari (UNMdP) for their cooperation.
This study was financially supported by the PICT 15 No. 0717 (ANPCyT, Argentina) and EXA 769/16 and EXA 871/18 (Universidad Nacional de Mar del Plata, Argentina).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Animal procedures and management protocols were approved by the Institutional Animal Care and Use Committee (RD 148/15) of the Faculty of Exact and Natural Sciences, National University of Mar del Plata, Mar del Plata, Argentina and carried out in accordance with the revised form of The Guide for the Care and Use of Laboratory Animals (National Research Council US 2011). Unnecessary animal suffering was avoided throughout the study.
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