Evaluation of a virus concentration method based on ultrafiltration and wet foam elution for studying viruses from large-volume water samples

Eva Forés; Marta Rusiñol; Marta Itarte; Sandra Martínez-Puchol; Miquel Calvo; Sílvia Bofill-Mas

doi:10.1016/j.scitotenv.2022.154431

Evaluation of a virus concentration method based on ultrafiltration and wet foam elution for studying viruses from large-volume water samples

Sci Total Environ. 2022 Jul 10:829:154431. doi: 10.1016/j.scitotenv.2022.154431. Epub 2022 Mar 9.

Authors

Eva Forés¹, Marta Rusiñol², Marta Itarte¹, Sandra Martínez-Puchol¹, Miquel Calvo³, Sílvia Bofill-Mas¹

Affiliations

¹ Laboratory of Viruses Contaminants of Water and Food, Genetics, Microbiology & Statistics Department at the University of Barcelona (UB), Barcelona, Catalonia, Spain; The Water Research Institute (IdRA), Universitat de Barcelona, Barcelona, Catalonia, Spain.
² Laboratory of Viruses Contaminants of Water and Food, Genetics, Microbiology & Statistics Department at the University of Barcelona (UB), Barcelona, Catalonia, Spain; Institute of Environmental Assessment & Water Research (IDAEA), CSIC, Barcelona, Catalonia, Spain. Electronic address: mrusinol@ub.edu.
³ Laboratory of Viruses Contaminants of Water and Food, Genetics, Microbiology & Statistics Department at the University of Barcelona (UB), Barcelona, Catalonia, Spain.

PMID: 35278558
DOI: 10.1016/j.scitotenv.2022.154431

Abstract

Assessing the presence of viruses in large-volume samples involves cumbersome methods that require specialized training and laboratory equipment. In this study, a large volume concentration (LVC) method, based on dead-end ultrafiltration (DEUF) and Wet Foam Elution™ technology, was evaluated in different type of waters and different microorganisms. Its recovery efficiency was evaluated through different techniques (infectivity assays and molecular detection) by spiking different viral surrogates (bacteriophages PhiX174 and MS2 and Coxsackie virus B5 (CVB5) and Escherichia coli (E. coli). Furthermore, the application of a secondary concentration step was evaluated and compared with skimmed milk flocculation. Viruses present in river water, seawater and groundwater samples were concentrated by applying LVC method and a centrifugal ultrafiltration device (CeUF), as a secondary concentration step and quantified with specific qPCR Human adenoviruses (HAdV) and noroviruses (NoVs). MS2 was used as process control, obtaining a mean viral recovery of 22.0 ± 12.47%. The presence of other viruses was also characterized by applying two different next-generation sequencing approaches. LVC coupled to a secondary concentration step based on CeUF allowed to detect naturally occurring viruses such as HAdV and NoVs in different water matrices. Using HAdV as a human fecal indicator, the highest viral pollution was found in river water samples (100% of positive samples), followed by seawater (83.33%) and groundwater samples (66.67%). The LVC method has also proven to be useful as a virus concentration method in the filed since HAdV and NoVs were detected in the river water and groundwater samples concentrated in the field. All in all, LVC method presents high concentration factor and a low limit of detection and provides viral concentrates useful for subsequent molecular analysis such as PCR and massive sequencing.

Keywords: Freshwater; Large volume concentration; Sea water; Ultrafiltration; Viral detection; Viral metagenomics.

MeSH terms

Adenoviruses, Human*
Escherichia coli
Humans
Norovirus*
Real-Time Polymerase Chain Reaction
Ultrafiltration
Water
Water Microbiology

Substances

Water