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The Optimization of Methods for the Collection of Aerosolized Murine Norovirus

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Abstract

Globally, norovirus is the most common gastroenteritis causing pathogen. Annually, norovirus causes 685 million cases of acute gastroenteritis and 200,000 deaths, worldwide. Recent evidence has suggested that norovirus can also be spread via aerosolization; however, an indoor generation source has yet to be determined. We optimized a sampling method for the collection of aerosolized norovirus using murine norovirus (MNV) as a surrogate. Optimization of the sampling method was performed using two bioaerosol samplers (SKC BioSampler and the NIOSH Bioaerosol Cyclone Sampler 251) and two sampling media (Hanks Balanced Salt Solution [HBSS] and Phosphate Buffered Saline [PBS]). Murine norovirus was aerosolized in a bioaerosol chamber and later collected using each sampler/media combination. Collected MNV was quantified using quantitative polymerase chain reaction (qPCR). Intact capsids of MNV were assessed using propidium monoazide dye in combination with qPCR and confirmed with transmission electron microscopy. Ten trials were conducted, with each trial lasting for 30 min. The SKC BioSampler collected a significantly higher concentration of MNV than the NIOSH-251 sampler did (p-value < 0.0001). However, there were no significant differences in the relative percent of MNV that remained viable between both samplers (p-value = 0.2215). The use of HBSS sampling media yielded a higher concentration of MNV than PBS media (p-value = 0.0125). However, PBS media maintained viability at a significantly higher percentage than HBSS media (p-value < 0.0001). The results support the optimization of a sampling method for the collection of aerosolized MNV and possibly norovirus in different sampling environments.

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Acknowledgements

The authors thank Dr. Skip Virgin’s Laboratory at Washington University School of Medicine for graciously donating the required amount of MNV. We acknowledge the National Institute for Occupational Safety and Health for allowing us to use their NIOSH Bioaerosol Cyclone samplers. We would also like the thank Dr. Tom Peters laboratory at the University of Iowa for technical support involving the creation and testing of the bioaerosol chamber. Lastly, we acknowledge Dr. Matthew Nonnenmann’s Lav Lab, including support staff, for laboratory assistance during this project.

Funding

This research was supported by a pilot project research training grant from the Heartland Center for Occupational Health and Safety at the University of Iowa. The Heartland Center is supported by Training Grant No. T42OH008491 from the Centers for Disease Control and Prevention/National Institute for Occupational Safety and Health. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

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Boles, C., Brown, G., Park, J.H. et al. The Optimization of Methods for the Collection of Aerosolized Murine Norovirus. Food Environ Virol 12, 199–208 (2020). https://doi.org/10.1007/s12560-020-09430-4

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