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Published (Version of Record)CC BY V4.0, Open Access
Journal article
Wastewater sample storage for physicochemical and microbiological analysis
Journal of virological methods, Vol.332, 115063
2025
Abstract
Highlights
• Most wastewater markers show no significant difference between 4°C and −80°C storage.
• Wastewater storage at 4°C maintains sample integrity for most markers for up to 12 weeks.
• Freeze-thaw cycles significantly degrade RNA viruses like SARS-CoV-2 in wastewater.
• CrAssphage DNA remains stable in frozen wastewater for up to one year of storage.
• Extractable RNA from wastewater increases in frozen samples, due to bacterial lysis.
Abstract
Wastewater-based epidemiology (WBE) is a crucial tool for health and environmental monitoring, providing real-time data on public health indicators by analysis of sewage samples. Ensuring the integrity of these samples from collection to analysis is paramount. This study investigates the effects of different cold-storage conditions on the integrity of wastewater samples, focusing on both microbiological markers (such as extractable nucleic acids, SARS-CoV-2, and crAssphage) and physicochemical parameters (including ammonium, orthophosphate, pH, conductivity, and turbidity). Composite samples from the combined raw wastewater influent from five wastewater treatment works in South Wales, UK, were stored at 4°C, −20°C, and −80°C, and subjected to up to six freeze-thaw cycles over one year. The study found significant effects of storage temperature on the preservation of certain WBE markers, with the best yield most frequently seen in samples stored at −80°C. However, the majority of WBE markers showed no significant difference between storage at −80°C or at 4°C, demonstrating that it may not always be necessary to archive wastewater samples at ultra-low temperatures, thus reducing CO2 emissions and laboratory energy costs. These findings underscore the importance of optimized storage conditions to maintain sample integrity, while ensuring accurate and reliable WBE data for public health and environmental monitoring.
Details
- Title
- Wastewater sample storage for physicochemical and microbiological analysis
- Authors/Creators
- Gordon Webster - Cardiff UniversityShrinivas Nivrutti Dighe - University Hospital of WalesWilliam B. Perry - Cardiff UniversityEwan H. Stenhouse - Cardiff UniversityDavey L. Jones - Bangor UniversityPeter Kille - Cardiff UniversityAndrew J. Weightman - Cardiff University
- Publication Details
- Journal of virological methods, Vol.332, 115063
- Publisher
- Elsevier B.V
- Number of pages
- 13
- Grant note
- Welsh Government under the Welsh National Wastewater Monitoring Programme: C035/2021/2022
This work was funded by the Welsh Government under the Welsh National Wastewater Monitoring Programme (C035/2021/2022).
- Identifiers
- 991005719239907891
- Copyright
- © 2024 The Author(s).
- Murdoch Affiliation
- Centre for Sustainable Farming Systems
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- Citation topics
- 1 Clinical & Life Sciences
- 1.81 Reproductive Biology
- 1.81.2160 Freeze Concentration
- Web Of Science research areas
- Biochemical Research Methods
- Biotechnology & Applied Microbiology
- Virology
- ESI research areas
- Microbiology