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Comparison of Skimmed Milk and Lanthanum Flocculation for Concentration of Pathogenic Viruses in Water

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Abstract

Concentration of viruses in water is necessary for detection and quantification of the viruses present, in order to evaluate microbiological barriers in water treatment plants and detect pathogenic viruses during waterborne outbreaks, but there is currently no standardised procedure. In this study, we implemented a previously described fast and simple lanthanum-based protocol for concentration of norovirus genogroup I (GI), genogroup II (GII) and hepatitis A virus (HAV) in drinking and surface water. We compared the results with those of a widely used skimmed milk flocculation method, followed by nucleic acid extraction and RT-qPCR detection. Three seeding levels, with intended concentrations 5 × 103, 5 × 104 and 5 × 105 genome copies/10 L, were added to drinking water or surface water. All seed levels were detected with both flocculation methods. Samples extracted with skimmed milk flocculation had on average 1.82, 1.86 and 1.38 times higher measured concentration of norovirus GI, GII and HAV, respectively, than those extracted with lanthanum flocculation, across all seeding levels and water types tested. Mengovirus was used as a positive process control. Mengovirus recovery was higher for skimmed milk (40.7% in drinking water, 26.0% in surface water) than for lanthanum flocculation (24.4% in drinking water, 9.7% in surface water). Together, this indicates that skimmed milk flocculation provides higher viral recovery than lanthanum flocculation. However, lanthanum-based flocculation can be performed much faster than skimmed milk flocculation (1.5 h versus 16 h flocculation time) and thus could be a good alternative for rapid monitoring of viruses in water.

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Acknowledgements

This study was funded by the Swedish Civil Contingencies Agency, within the projects “Ökad förmåga till detektion av virus i livsmedel och humanprover för förbättrad smittspårning”, and “Stärkt beredskapskapacitet via rationell laboratoriediagnostik samt förenklad provberedning, pre-PCR processing”. The authors would like to thank Norrvatten (Görväln vattenverk) for surface water samples, Kåre Bondesson at the University Hospital in Uppsala for providing positive norovirus stool samples, and Anna Charlotte Schultz at Technical University of Denmark for cell culture supernatant from mengovirus.

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  1. Emmy Borgmästars

    Present address: Department of Surgical and Perioperative Sciences, Surgery, Umeå University, Umeå, Sweden

Authors and Affiliations

  1. Science Division, Biology Department, Swedish Food Agency, Hamnesplanaden 5, 75319, Uppsala, Sweden

    Emmy Borgmästars, Sofia Persson, Maria Hellmér, Magnus Simonsson & Ronnie Eriksson

  2. European Union Reference Laboratory (EURL) for Foodborne Viruses, Uppsala, Sweden

    Sofia Persson, Maria Hellmér, Magnus Simonsson & Ronnie Eriksson

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  1. Emmy Borgmästars
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Correspondence to Emmy Borgmästars or Ronnie Eriksson.

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Stool samples used in this study have been de-identified and thus no ethical approval was needed according to the Swedish Act concerning the Ethical Review of Research Involving Humans (2003:460).

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Borgmästars, E., Persson, S., Hellmér, M. et al. Comparison of Skimmed Milk and Lanthanum Flocculation for Concentration of Pathogenic Viruses in Water. Food Environ Virol 13, 380–389 (2021). https://doi.org/10.1007/s12560-021-09477-x

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