Characterization of the efficiency and uncertainty of skimmed milk flocculation for the simultaneous concentration and quantification of water-borne viruses, bacteria and protozoa

Eloy Gonzales-Gustavson, Yexenia Cárdenas-Youngs, Miquel Calvo, Marcelle Figueira Marques da Silva, Ayalkibet Hundesa, Inmaculada Amorós, Yolanda Moreno, Laura Moreno-Mesonero, Rosa Rosell, Llilianne Ganges, Rosa Araujo, Rosina Girones

Research output: Contribution to journalArticlepeer-review

14 Scopus citations


In this study, the use of skimmed milk flocculation (SMF) to simultaneously concentrate viruses, bacteria and protozoa was evaluated. We selected strains of faecal indicator bacteria and pathogens, such as Escherichia coli and Helicobacter pylori. The viruses selected were adenovirus (HAdV 35), rotavirus (RoV SA-11), the bacteriophage MS2 and bovine viral diarrhoea virus (BVDV). The protozoa tested were Acanthamoeba, Giardia and Cryptosporidium. The mean recoveries with q(RT)PCR were 66% (HAdV 35), 24% (MS2), 28% (RoV SA-11), 15% (BVDV), 60% (E. coli), 30% (H. pylori) and 21% (Acanthamoeba castellanii). When testing the infectivity, the mean recoveries were 59% (HAdV 35), 12% (MS2), 26% (RoV SA-11) and 0.7% (BVDV). The protozoa Giardia lamblia and Cryptosporidium parvum were studied by immunofluorescence with recoveries of 18% and 13%, respectively. Although q(RT)PCR consistently showed higher quantification values (as expected), q(RT)PCR and the infectivity assays showed similar recoveries for HAdV 35 and RoV SA-11. Additionally, we investigated modelling the variability and uncertainty of the recovery with this method to extrapolate the quantification obtained by q(RT)PCR and estimate the real concentration. The 95% prediction intervals of the real concentration of the microorganisms inoculated were calculated using a general non-parametric bootstrap procedure adapted in our context to estimate the technical error of the measurements. SMF shows recoveries with a low variability that permits the use of a mathematical approximation to predict the concentration of the pathogen and indicator with acceptable low intervals. The values of uncertainty may be used for a quantitative microbial risk analysis or diagnostic purposes.

Original languageEnglish
Pages (from-to)46-53
Number of pages8
JournalJournal of Microbiological Methods
StatePublished - 1 Mar 2017

Bibliographical note

Funding Information:
This study was funded by the Water Challenges for a Changing World Joint Programming Initiative (W2013-095-C03-01), the Spanish Ministry of Economy and Competitiveness (MINECO ? AGL2014-55081-R) and the Grup de Recerca Consolidat: Virus, bacteris i protozous d'inter?s en salut p?blica, aigua y aliments (Generalitat de Catalunya, Virbap ? 2014-SGR-914).

Publisher Copyright:
© 2017 The Author(s)

Copyright 2018 Elsevier B.V., All rights reserved.


  • Microorganisms
  • Real concentration
  • Recovery
  • Skimmed milk flocculation
  • Water contamination
  • q(RT)PCR

Fingerprint Dive into the research topics of 'Characterization of the efficiency and uncertainty of skimmed milk flocculation for the simultaneous concentration and quantification of water-borne viruses, bacteria and protozoa'. Together they form a unique fingerprint.

Cite this