Simultaneous Concentration of Enteric Viruses and Protozoan Parasites: A Protocol Based on Tangential Flow Filtration and Adapted to Large Volumes of Surface and Drinking Waters |
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Authors: | Skraber Sylvain Gantzer Christophe Helmi Karim Hoffmann Lucien Cauchie Henry-Michel |
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Institution: | (1) Department of Environment and Agro-biotechnologies (EVA), Centre de Recherche Public-Gabriel Lippmann, 41 rue du Brill, L-4422 Belvaux, Luxembourg;(2) Laboratoire de Chimie Physique et Microbiologie pour l’Environnement (LCPME), Faculté de Pharmacie, UMR 7564, CNRS/Nancy Université, 5 rue Albert Lebrun, F-54000 Nancy, France |
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Abstract: | A filtration system, based on tangential flow filtration (TFF) followed by ultracentrifugation was developed in order to concentrate
simultaneously viruses and parasites from large volumes of water. For TFF, no pre-treatment of the membrane is performed but
a post-rinsing step using high pH-beef extract-based eluant. Applying our protocol to 20 l of surface waters spiked with vaccinal
poliovirus-1, ϕX174 and MS2 bacteriophages resulted in an averaged viral recovery of 75% by TFF and 91% by ultracentrifugation
(total viral recovery of 70%). Our protocol was further applied to 31 environmental samples including surface, ground and
drinking waters from the Grand Duchy of Luxembourg in order to assess the occurrence of protozoan parasites (Cryptosporidium parvum and Giardia lamblia (oo)cysts), pathogenic viruses (enterovirus, norovirus and adenovirus) and infectious bacteriophages (somatic coliphages
and F-specific phages) in these samples. High viral recovery rates of > 70% were confirmed concentrating environmental strains
of somatic and F-specific coliphages from non-spiked surface waters. Parasites and enteric viruses were detected in 86 and
40% of the surface waters used for drinking water production, respectively. Infectious bacteriophages were isolated from all
surface waters and in two out of seven (29%) groundwaters revealing a susceptibility of the corresponding wells to viral pollution.
TFF-based method proved to be efficient for surveying the occurrence of non-bacterial pathogens such as enteric viruses and
protozoan parasites in large volumes of environmental waters. |
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Keywords: | Virus concentration Tangential flow filtration Ultracentrifugation Drinking water Surface water |
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