Effects of freezing and storage temperature on MS2 viability

Matthew R. Olson, Richard P Axler, Randall E Hicks

Research output: Contribution to journalArticlepeer-review

61 Scopus citations


Monitoring human enteric virus levels in domestic wastewater effluent is crucial to protecting human health. Occasionally, during intensive sampling, wastewater samples must be stored for later viral analysis. Little data exist regarding how enteric viruses survive during storage at different temperatures in secondary treated wastewater. During a field-scale study assessing pathogen removal performance by various onsite treatment technologies, the MS2 bacteriophage, an indicator of enteric viruses, was inoculated into septic tank (STE), sand filter, peat filter and constructed wetland (CW) effluents to determine virus decay at various storage temperatures. Virus stored at temperatures ≥10°C and at -20°C decayed nearly twice as fast as those stored at 4°C or -80°C. Decreased water quality decreased viral decay rates at 4°C and -80°C, with slowest decay occurring in STE and the fastest in sterile PBS and low pH peat effluent. In CW effluent after 8 days, less MS2 was inactivated when stored at 4°C (20%) compared to -80°C (58%); however, during extended storage (∼300 days), less MS2 was inactivated at -80°C (75%) compared) to 4°C (93%). We recommend that viruses in wastewater be stored in the dark at 4°C unless storage for >40 days is necessary.

Original languageEnglish (US)
Pages (from-to)147-152
Number of pages6
JournalJournal of Virological Methods
Issue number2
StatePublished - Dec 15 2004

Bibliographical note

Funding Information:
We thank D. Battegelli for providing the MS2 virus and E. coli host, and R. Karim for assistance in experimental design and techniques. The Legislative Commission on Minnesota Resources, Minnesota Sea Grant (supported by the National Oceanic and Atmospheric Administration of Sea Grant, United States Department of Commerce under Grant No. USDOC/NA46RG0101, Project No. R/A-11 to RPA), and the Natural Resource Research Institute provided funding and technical support for this project. Summer fellowships were provided by the University of Minnesota-Duluth Graduate Biology Program and the University of Minnesota Water Resources Science Graduate Program. This paper is journal reprint number JR 504 of the Minnesota Sea Grant College Program and contribution # 367 from the NRRI Center for Water and the Environment.

Copyright 2012 Elsevier B.V., All rights reserved.


  • Enteric viruses
  • MS2 bacteriophage
  • Onsite treatment
  • Virus storage
  • Wastewater pathogens


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