Determination of Giardia cyst viability in environmental and faecal samples by immunofluorescence, fluorogenic dye staining and differential interference contrast microscopy

Letters in Applied Microbiology
L ThiriatJanine Schwartzbrod

Abstract

Amongst the techniques suggested for the determination of Giardia cyst viability, the use of the fluorogenic dyes, fluorescein diacetate (FDA) and propidium iodide (PI) is the most often recommended, even though it appears to overestimate the number of viable cysts. In the present study, the replacement of FDA with 4',6-diamidino-2-phenylindole (DAPI) allowed simultaneous direct immunofluorescence with monoclonal antibody labelled with fluorescein isothiocyanate (MAb-FITC). Under these conditions, it was possible both to quantify the cysts according to the immunofluorescence technique, and to appreciate their viability by using fluorogenic dye staining (DAPI and PI) and differential interference contrast (DIC) microscopy. This method proved to be significantly better than the counting methods normally suggested. The technique has been applied to Giardia cysts recovered from faeces and wastewater sludge.

Citations

Mar 9, 2004·Journal of Microbiological Methods·Isabelle BertrandJanine Schwartzbrod
Jan 9, 2001·Clinical Microbiology Reviews·T B Gardner, D R Hill
Nov 6, 2013·Biotechnology Advances·Anson V KoehlerRobin B Gasser
Jun 20, 2006·Environmental Research·P KaranisK Stojanova
Sep 10, 2011·Veterinary Parasitology·G H GritT Geurden
Nov 19, 2013·Journal of Microbiological Methods·Lucy J RobertsonEdwin Claerebout
Feb 20, 2010·PLoS Neglected Tropical Diseases·Hugo Aguilar-DíazJulio C Carrero
Jun 27, 2017·Water Research·Artemis EfstratiouPanagiotis Karanis

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