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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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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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Acknowledgements

This work was funded by the FNR (National Research Fund) from Luxembourg (SECAL Program, KAWA Project, FNR/03/07/07). The authors thank Laurent Diez, Nicolas Bonjean, Delphine Collard and Céline Krzykala for their excellent technical assistance and Jean Louis Belot and Daniel Perez from SARTORIUS Corporation for their support. The authors also thank Jean-Paul Lickes, Georges Kraus, Isabelle Kolber, Gerard Zimmer, Claude Mathieu and Norbert Entringer for their kind cooperation at the different sampling sites. The authors acknowledge Ana Maria de Roda Husman and Lodder Willemijn from the RIVM institute (Bilthoven, The Netherlands) for having provided the different norovirus strains that have been used as positive controls in this study.

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  1. Department of Environment and Agro-biotechnologies (EVA), Centre de Recherche Public-Gabriel Lippmann, 41 rue du Brill, L-4422, Belvaux, Luxembourg

    Skraber Sylvain, Helmi Karim, Hoffmann Lucien & Cauchie Henry-Michel

  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

    Gantzer Christophe

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  1. Skraber Sylvain
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Correspondence to Skraber Sylvain.

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Sylvain, S., Christophe, G., Karim, H. et al. 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. Food Environ Virol 1, 66–76 (2009). https://doi.org/10.1007/s12560-009-9011-z

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  • DOI: https://doi.org/10.1007/s12560-009-9011-z

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