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Type: Artigo
Title: Fluorescence decay of dyed protozoa: differences between stressed and non‐stressed cysts
Author: Santos, Samuel Ricardo dos
Branco, Nilson
Franco, Regina Maura Bueno
Paterniani, José Euclides Stipp
Katsumata, Masakazu
Barlow, Peter W.
Gallep, Cristiano de Mello
Abstract: Several series of tests have shown that fresh, intact samples of Giardia duodenalis and Cryptosporidium parvum (oo) cysts are not marked by fluorescent probes such as carboxyfluorcein-succinimidyl-diacetate-ester (CFDA-SE), C12-resazurin and SYTOX (R) Green, probably because of their robust cell walls. These dyes fail to indicate the viability of such protozoa and allow negative responses to be recorded from living and infectious samples. Cryptosporidium parvum showed stronger isolation from chemicals, with living oocysts remaining unstained by the probe for up to 90 days after extraction. However, in further fluorescence decay (FD) experiments run with G. duodenalis samples stained using CFDA-SE (comprising living, non-stressed but aged cysts, heat-killed samples and UV-C-stressed samples) each showed a different FD decay profile, here studied in seven series of tests of five replicates each. The FD profiles were fitted by double-exponential decay kinetics, with the decay constant k(2) being five times higher than k(1). This FD procedure is fast and can be easily reproduced in 10 steps, taking similar to 1 h of laboratory work for already purified samples. Copyright (C) 2015 John Wiley & Sons, Ltd.
Several series of tests have shown that fresh, intact samples of Giardia duodenalis and Cryptosporidium parvum (oo)cysts are not marked by fluorescent probes such as carboxyfluorcein‐succinimidyl‐diacetate‐ester (CFDA‐SE), C12‐resazurin and SYTOX® Green,
Subject: Microscopia de fluorescência
Giardia lamblia
Cryptosporidium parvum
Country: Reino Unido
Editor: John Wiley & Sons
Citation: Fluorescence Decay Of Dyed Protozoa: Differences Between Stressed And Non-stressed Cysts. Wiley-blackwell, v. 30, p. 1139-1147 NOV-2015.
Rights: fechado
Identifier DOI: 10.1002/bio.2872
Date Issue: 2015
Appears in Collections:FEAGRI - Artigos e Outros Documentos
IB - Artigos e Outros Documentos
FT - Artigos e Outros Documentos

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